Substituted 6,5-fused bicyclic heteroaryl compounds

ABSTRACT

The present invention relates to substituted 6,5-fused bicyclic heteroaryl compounds. The present invention also relates to pharmaceutical compositions containing these compounds and methods of treating cancer by administering these compounds and pharmaceutical compositions to subjects in need thereof.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/536,162, filed Nov. 7, 2014 (to issue as U.S. Pat. No. 9,637,472 onMay 2, 2017), which is a continuation of U.S. application Ser. No.14/070,675, filed Nov. 4, 2013 (now U.S. Pat. No. 8,962,620), which is acontinuation of U.S. application Ser. No. 13/938,067, filed Jul. 9, 2013(now U.S. Pat. No. 8,598,167), which is a continuation of InternationalApplication PCT/US2012/026953, with an international filing date of Feb.28, 2012, which claims priority to, and the benefit of, U.S. provisionalapplication Nos. 61/447,632, filed Feb. 28, 2011, 61/447,625, filed Feb.28, 2011, 61/447,626 filed Feb. 28, 2011, 61/447,627 filed Feb. 28,2011, and 61/447,629 filed Feb. 28, 2011, the entire contents of each ofwhich are incorporated herein by reference in their entireties.

INCORPORATION-BY-REFERENCE OF SEQUENCE LISTING

The contents of the text file named “41478503001WOST25.txt”, which wascreated on Feb. 28, 2012 and is 2 KB in size, are hereby incorporated byreference in their entireties.

BACKGROUND OF THE INVENTION

In eukaryotic cells DNA is packaged with histones to form chromatin.Changes in the ordered structure of chromatin can lead to alterations intranscription of associated genes. Control of changes in chromatinstructure (and hence of transcription) is mediated by covalentmodifications to histones, most notably of their N-terminal tails. Thesemodifications are often referred to as epigenetic because they can leadto heritable changes in gene expression, but do not affect the sequenceof the DNA itself. Covalent modifications (for example, methylation,acetylation, phosphorylation, and ubiquitination) of the side chains ofamino acids are enzymatically mediated. The selective addition of methylgroups to specific amino acid sites on histones is controlled by theaction of a unique family of enzymes known as histone methyltransferases(HMTs).

The orchestrated collection of biochemical systems behindtranscriptional regulation must be tightly controlled in order for cellgrowth and differentiation to proceed optimally. Disease states resultwhen these controls are disrupted by aberrant expression and/or activityof the enzymes responsible for DNA and histone modification. In humancancers, for example, there is a growing body of evidence to suggestthat dysregulated epigenetic enzyme activity contributes to theuncontrolled cell proliferation associated with cancer as well as othercancer-relevant phenotypes such as enhanced cell migration and invasion.Beyond cancer, there is growing evidence for a role of epigeneticenzymes in a number of other human diseases, including metabolicdiseases (such as diabetes), inflammatory diseases (such as Crohn'sdisease), neurodegenerative diseases (such as Alzheimer's disease) andcardiovascular diseases. Therefore, selectively modulating the aberrantaction of epigenetic enzymes holds great promise for the treatment of arange of diseases.

Polycomb group (PcG) and trithorax group (trxG) proteins are known to bepart of the cellular memory system. See, e.g., Francis et al. (2001) NatRev Mol Cell Biol 2:409-21 and Simon et al. (2002) Curr Opin Genet Dev12:210-8. In general, PcG proteins are transcriptional repressors thatmaintain the “off state,” and trxG proteins are transcriptionalactivators that maintain the “on state.” Because members of PcG and trxGproteins contain intrinsic histone methyltransferase (HMTase) activity,PcG and trxG proteins may participate in cellular memory throughmethylation of core histones. See, e.g., Beisel et al. (2002) Nature419:857-62; Cao et al. (2002) Science 298:1039-43; Czermin et al. (2002)Cell 111:185-96; Kuzmichev et al. (2002) Genes Dev 16:2893-905; Milne etal. (2002) Mol Cell 10:1107-17; Muller et al. (2002) Cell 111:197-208;and Nakamura et al. (2002) Mol Cell 10:1119-28.

Biochemical and genetic studies have provided evidence that DrosophilaPcG proteins function in at least two distinct protein complexes, thePolycomb repressive complex 1 (PRC1) and the ESC-E(Z) complex (alsoknown as Polycomb repressive complex 2 (PRC2)). Otte et al. (2003) CurrOpin Genet Dev 13:448-54. Studies in Drosophila have demonstrated thatthe ESC-E(Z)/EED-EZH2 (i.e., PRC2) complexes have intrinsic histonemethyltransferase activity. Although the compositions of the complexesisolated by different groups are slightly different, they generallycontain EED, EZH2, SUZ12, and RbAp48 or Drosophila homologs thereof.However, a reconstituted complex comprising only EED, EZH2, and SUZ12retains histone methyltransferase activity for lysine 27 of histone H3.U.S. Pat. No. 7,563,589.

Of the various proteins making up PRC2 complexes, EZH2 (Enhancer ofZeste Homolog 2) is the catalytic subunit. The catalytic site of EZH2 inturn is present within a SET domain, a highly conserved sequence motif(named after Su(var)3-9, Enhancer of Zeste, Trithorax) that is found inseveral chromatin-associated proteins, including members of both theTrithorax group and Polycomb group. SET domain is characteristic of allknown histone lysine methyltransferases except the H3-K79methyltransferase DOT1.

In addition to Hox gene silencing, PRC2-mediated histone H3-K27methylation has been shown to participate in X-inactivation. Plath etal. (2003) Science 300:131-5; Silva et al. (2003) Dev Cell 4:481-95.Recruitment of the PRC2 complex to Xi and subsequent trimethylation onhistone H3-K27 occurs during the initiation stage of X-inactivation andis dependent on Xist RNA. Furthermore, EZH2 and its associated histoneH3-K27 methyltransferase activity were found to mark differentially thepluripotent epiblast cells and the differentiated trophectoderm, andconsistent with a role of EZH2 in maintaining the epigeneticmodification patterns of pluripotent epiblast cells, Cre-mediateddeletion of EZH2 results in loss of histone H3-K27 methylation in thecells. Erhardt et al. (2003) Development 130:4235-48). Further, studiesin prostate and breast cancer cell lines and tissues have revealed astrong correlation between the levels of EZH2 and SUZ12 and theinvasiveness of these cancers, indicating that dysfunction of the PRC2complex may contribute to cancer. Bracken et al. (2003) EMBOJ22:5323-35; Kirmizis et al. (2003) Mol Cancer Ther 2:113-21; Kleer etal. (2003) Proc Natl Acad Sci USA 100:11606-11; Varambally et al. (2002)Nature 419:624-9.

Recently, somatic mutations of tyrosine 641 (Y641C, Y641F, Y641N, Y641Sand Y641H; sometimes also referred to as Y646C, Y646F, Y646N, Y646S andY646H, respectively) of EZH2 were reported to be associated withfollicular lymphoma (FL) and the germinal center B cell-like (GCB)subtype of diffuse large B-cell lymphoma (DLBCL). Morin et al. (2010)Nat Genet 42:181-5. In all cases, occurrence of the mutant EZH2 gene wasfound to be heterozygous, and expression of both wild-type and mutantalleles was detected in the mutant samples profiled by transcriptomesequencing. It was also demonstrated that all of the mutant forms ofEZH2 could be incorporated into the multi-protein PRC2 complex, but thatthe resulting complexes lacked the ability to catalyze methylation ofthe H3-K27 equivalent residue of a peptidic substrate. Hence, it wasconcluded that the disease-associated changes at Tyr641 of EZH2 resultedin loss of function with respect to EZH2-catalyzed H3-K27 methylation.

SUMMARY OF THE INVENTION

In one aspect, the present invention features substituted 6,5-fusedbicyclic heteroaryl compound of Formula (I) below or a pharmaceuticallyacceptable salt or ester thereof

In this formula:

-   -   X₁ is NR₇ or CR₇;    -   X₂ is N, NR₈, CR₈, O, or S;    -   X₃ is NR₈, CR₈, O, or S;    -   X₄ is C or N;    -   Y₁ is N or CH;    -   Y₂ is N or CR₆;    -   Y₃ is N, or CR₁₁, and at least one of X₁, X₂, X₃, X₄, Y₁, Y₂,        and Y₃ is N or NR₇;    -   each of R₁, R₅, R₉, and R₁₀, independently, is H or C₁-C₆ alkyl        optionally substituted with one or more substituents selected        from the group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆        alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, and C₃-C₈ cycloalkyl;    -   each of R₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₁ is H, halo, hydroxyl,        COOH, cyano, or R_(S1), in which R_(S1) is C₁-C₃ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl, C(O)O—C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and R_(S1) is optionally substituted with one or        more substituents selected from the group consisting of halo,        hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl,        amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5        to 6-membered heteroaryl;    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which each of        R_(a) and R_(b), independently is H or R_(S3) and each of R_(S2)        and R_(S3), independently is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl; or R_(a) and        R_(b), together with the N atom to which they are attached, form        a 4 to 7-membered heterocycloalkyl ring having 0 or 1 additional        heteroatoms to the N atom; and each of R_(S2), R_(S3), and the 4        to 7-membered heterocycloalkyl ring containing R_(a) and R_(b),        is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is        a bond or C₁-C₃ alkyl linker each optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano,        —OR_(c), —NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c),        —C(O)OR_(c), —C(O)NR_(c)R_(d), —NR_(d)C(O)R_(c),        —NR_(d)C(O)OR_(c), —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in        which each of R_(c) R_(d), and R_(d′), independently is H or        R_(S5), A⁻ is a pharmaceutically acceptable anion, each of        R_(S4) and R_(S5), independently, is C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, or 5        to 6-membered heteroaryl, or R_(c) and R_(d), together with the        N atom to which they are attached, form a 4 to 7-membered        heterocycloalkyl ring having 0 or 1 additional heteroatoms to        the N atom, and each of R_(S4), R_(S5), and the 4 to 7-membered        heterocycloalkyl ring containing R_(c) and R_(d), is optionally        substituted with one or more -Q₃-T₃, wherein Q₃ is a bond or        C₁-C₃ alkyl linker each optionally substituted with halo, cyano,        hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from the group        consisting of H, halo, cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, 5 to 6-membered        heteroaryl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and        —C(O)NR_(e)R_(f), each of R_(e) and R_(f) independently being H        or C₁-C₆ alkyl; or -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two        neighboring -Q₂-T₂, together with the atoms to which they are        attached form a 5- or 6-membered ring optionally containing 1-4        heteroatoms selected from N, O and S and optionally substituted        with one or more substituents selected from the group consisting        of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl,        and 5 to 6-membered heteroaryl; provided that -Q₂-T₂ is not H;    -   each R₇ independently is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄        alkyl linker, or C₂-C₄ alkenyl linker, each linker optionally        substituted with halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄        is H, halo, cyano, NR_(g)R_(h), —OR_(g), —C(O)R_(g),        —C(O)OR_(g), —C(O)NR_(g)R_(h), —C(O)NR_(g)OR_(h),        —NR_(g)C(O)R_(h), —S(O)₂R_(g), or R_(S6), in which each of R_(g)        and R_(h), independently is H or R_(S7), each of R_(S6) and        R_(S7), independently is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and each of R_(S6) and R_(S7) is optionally        substituted with one or more -Q₅-T₅, wherein Q₅ is a bond, C(O),        or C₁-C₃ alkyl linker, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl and T₅ is        optionally substituted with one or more substituents selected        from the group consisting of halo, C₁-C₆ alkyl, hydroxyl, cyano,        C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, and 5 to 6-membered heteroaryl except when T₅        is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; provided that        -Q₄-T₄ is not H; and    -   each of R₈ and R₁₁, independently, is H, halo, hydroxyl, COOH,        cyano, R_(S8), OR_(S8), or COOR_(S8), in which R_(S8) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S8) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino;    -   provided that the compound is not    -   N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,5-dimethylthiophen-3-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-(2,3-dihydro-1,4-benzodioxin-6-yl)-N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1,3-dimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1,3,6-trimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-6-methyl-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,        or    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide.

In another aspect, the present invention features substituted 6,5-fusedbicyclic heteroaryl compound of Formula (I) above or a pharmaceuticallyacceptable salt or ester thereof, in which

-   -   X₁ is NR₇ or CR₇;    -   X₂ is N, NR₈, CR₈, O, or S;    -   X₃ is NR₈, CR₈, O, or S;    -   X₄ is C or N;    -   Y₁ is N or CH;    -   Y₂ is N or CR₆;    -   Y₃ is N, or CR₁₁, and at least one of X₁, X₂, X₃, X₄, Y₁, Y₂,        and Y₃ is N or NR₇;    -   each of R₁, R₅, R₉, and R₁₀, independently, is H or C₁-C₆ alkyl        optionally substituted with one or more substituents selected        from the group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆        alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, and C₃-C₈ cycloalkyl;    -   each of R₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₁ is H, halo, hydroxyl,        COOH, cyano, or R_(S1), in which R_(S1) is C₁-C₃ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl, C(O)O—C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and R_(S1) is optionally substituted with one or        more substituents selected from the group consisting of halo,        hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl,        amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5        to 6-membered heteroaryl;    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which R_(S2) is        C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl        containing at least one N or O atom, each of R_(a) and R_(b),        independently is H or R_(S3), and R_(S3), independently is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl; or R_(a) and R_(b), together with the N atom to        which they are attached, form a 4 to 7-membered heterocycloalkyl        ring having 0 or 1 additional heteroatoms to the N atom; and        each of R_(S2), R_(S3), and the 4 to 7-membered heterocycloalkyl        ring containing R_(a) and R_(b), is optionally substituted with        one or more -Q₂-T₂, wherein Q₂ is a bond or C₁-C₃ alkyl linker        each optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₂ is H, halo, cyano, —NR_(c)R_(d),        —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c), —C(O)OR_(c),        —C(O)NR_(c)R_(d), —NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c),        —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in which each of        R_(c) R_(d), and R_(d′), independently is H or R_(S5), A⁻ is a        pharmaceutically acceptable anion, each of R_(S4) and R_(S5),        independently, is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl,        or R_(c) and R_(d), together with the N atom to which they are        attached, form a 4 to 7-membered heterocycloalkyl ring having 0        or 1 additional heteroatoms to the N atom, and each of R_(S4),        R_(S5), and the 4 to 7-membered heterocycloalkyl ring containing        R_(c) and R_(d), is optionally substituted with one or more        -Q₃-T₃, wherein Q₃ is a bond or C₁-C₃ alkyl linker each        optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₃ is selected from the group consisting of halo,        cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        7-membered heterocycloalkyl, 5 to 6-membered heteroaryl, OR_(e),        COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and —C(O)NR_(e)R_(f), each        of R_(e) and R_(f) independently being H or C₁-C₆ alkyl; or        -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two neighboring -Q₂-T₂,        together with the atoms to which they are attached form a 5- or        6-membered ring optionally containing 1-4 heteroatoms selected        from N, O and S and optionally substituted with one or more        substituents selected from the group consisting of halo,        hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,        mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5 to        6-membered heteroaryl; provided that -Q₂-T₂ is not H;    -   each R₇ independently is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄        alkyl linker, or C₂-C₄ alkenyl linker, each linker optionally        substituted with halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄        is H, halo, cyano, NR_(g)R_(h), —OR_(g), —C(O)R_(g),        —C(O)OR_(g), —C(O)NR_(g)R_(h), —C(O)NR_(g)OR_(h),        —NR_(g)C(O)R_(h), —S(O)₂R_(g), or R_(S6), in which each of R_(g)        and R_(b), independently is H or R_(S7), each of R_(S6) and        R_(S7), independently is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and each of R_(S6) and R_(S7) is optionally        substituted with one or more -Q₅-T₅, wherein Q₅ is a bond, C(O),        or C₁-C₃ alkyl linker, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl and T₅ is        optionally substituted with one or more substituents selected        from the group consisting of halo, C₁-C₆ alkyl, hydroxyl, cyano,        C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, and 5 to 6-membered heteroaryl except when T₅        is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; provided that        -Q₄-T₄ is not H; and    -   each of R₈ and R₁₁, independently, is H, halo, hydroxyl, COOH,        cyano, R_(S8), OR_(S8), or COOR_(S8), in which R_(S8) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S8) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino.

In one subset of the compounds of Formula (I), at least one of Y₁, Y₃,and X₄ is N and when X₄ is C, Y₁ is N, Y₂ is CR₆, and Y₃ is CR₁₁, thenX₂ is CR₈.

One subset of the compounds of Formula (I) includes those of Formula(Ia):

Another subset of the compounds of Formula (I) includes those of Formula(Ib).

The compounds of Formulae (I), (Ia), and (Ib) can include one or more ofthe following features:

X₄ is C.

X₂ is N or CH.

X₃ is CR₈.

Y₃ is CR₁₁.

R₆ is phenyl substituted with one or more -Q₂-T₂.

R₆ is 5 to 6-membered heteroaryl containing 1-3 additional heteroatomsselected from N, O, and S and optionally substituted with one or more-Q₂-T₂, provided that the heteroaryl is not thiophenyl.

R₆ is pyridinyl, pyrazolyl, pyrimidinyl, or furyl, each of which isoptionally substituted with one or more -Q₂-T₂.

R₆ is halo, C₁-C₃ alkyl substituted with one or more -Q₂-T₂, C₂-C₆alkenyl, C₄-C₆ cycloalkyl, C(O)H, OR_(a), or —C(O)R_(a), in which R_(a)is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl.

R₆ is 4 to 7-membered heterocycloalkyl optionally substituted with oneor more -Q₂-T₂, in which -Q₂-T₂ is oxo or Q₂ is a bond and T₂ is—OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), —S(O)₂R_(c), C₁-C₆alkyl, or 4 to 7-membered heterocycloalkyl, each of which is optionallysubstituted with one or more -Q₃-T₃ when R_(c) or R_(d) is not H.

R₆ is piperidinyl, 2,2,6,6-tetramethyl-piperidinyl,1,2,3,6-tetrahydropyridinyl,2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, orpyrrolidinyl, each of which is optionally substituted with one or more-Q₂-T₂.

Q₃ is a bond or C₁-C₃ alkyl linker and T₃ is selected from the groupconsisting of C₁-C₃ alkyl, halo, OR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and—C(O)NR_(e)R_(f).

R₇ is C₁-C₆ alkyl, C₂-C₄ alkenyl, C₄-C₆ cycloalkyl, 4 to 7-memberedheterocycloalkyl, or C₆-C₁₀ aryl, each optionally substituted with oneor more -Q₅-T₅.

R₇ is cyclopentyl.

R₇ is unsubstituted C₁-C₆ alkyl.

R₇ is isopropyl.

R₇ is 5 to 6-membered heterocycloalkyl optionally substituted with oneor more -Q₅-T₅.

R₇ is piperidinyl optionally substituted with one -Q₅-T₅.

T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈ cycloalkyl, C₆-C₁₀aryl, or 4 to 7-membered heterocycloalkyl.

Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4 to 7-memberedheterocycloalkyl.

Q₅ is CO and T₅ is C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈ cycloalkyl, or 4 to7-membered heterocycloalkyl.

Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

R₁₁ is H.

Each of R₂ and R₄, independently is H or C₁-C₆ alkyl optionallysubstituted with amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, orC₆-C₁₀ aryl.

Each of R₂ and R₄, independently is C₁-C₃ alkyl optionally substitutedwith C₁-C₆ alkoxyl.

Each of R₂ and R₄ is methyl.

R₁ is H.

R₅ is H, methyl, or ethyl.

R₃ is H.

is selected from the group consisting of indolyl, isoindolyl,indolizinyl, benzofuryl, isobenzofuryl, benzo[b]thienyl, benzoxazolyl,benzthiazolyl, benzimidazolyl, benzotriazolyl, benzoxadiazolyl,benzothiadiazolyl, purinyl, indazolyl, pyrrolopyridinyl,imidazopyridinyl, pyrazolopyridinyl, pyrrolopyrazinyl, imidazopyrazinyl,pyrazolopyrazinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl,pyrrolopyridazinyl, imidazopyridazinyl, pyrazolopyridazinyl,furopyridinyl, thienopyridinyl, furopyrazinyl, thienopyrazinyl,oxazolopyridinyl, isoxazolopyridinyl, thiazolopyridinyl,isothiazolopyridinyl, oxadiazolopyridinyl, thiadiazolopyridinyl,triazolopyridinyl, oxazolopyrazinyl, isoxazolopyrazinyl,thiazolopyrazinyl, isothiazolopyrazinyl, oxadiazolopyrazinyl,thiadiazolopyrazinyl, triazolopyrazinyl, furopyrimidinyl,thienopyrimidinyl, furopyridazinyl, thienopyridazinyl,oxazolopyrimidinyl, isoxazolopyrimidinyl, thiazolopyrimidinyl,isothiazolopyrimidinyl, oxadiazolopyrimidinyl, thiadiazolopyrimidinyl,triazolopyrimidinyl, oxazolopyridazinyl, isoxazolopyridazinyl,thiazolopyridazinyl, isothiazolopyridazinyl, oxadiazolopyridazinyl,thiadiazolopyridazinyl, triazolopyridazinyl, and imidazotriazinyl.

is selected from the group consisting of

is selected from the group consisting of

is selected from the group consisting of

The present invention also provides pharmaceutical compositionscomprising one or more pharmaceutically acceptable carriers and one ormore compounds selected from those of Formulae (I), (Ia), and (Ib),N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,5-dimethylthiophen-3-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,6-(2,3-dihydro-1,4-benzodioxin-6-yl)-N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1,3-dimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1,3,6-trimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-6-methyl-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,and6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide.

Another aspect of this invention is a method of treating or preventingcancer. The method includes administering to a subject in need thereof atherapeutically effective amount of one or more compounds selected fromthose of Formulae (I), (Ia), and (Ib),N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,5-dimethylthiophen-3-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,6-(2,3-dihydro-1,4-benzodioxin-6-yl)-N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1,3-dimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1,3,6-trimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-6-methyl-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,and6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide.

For example, the method comprises the step of administering to a subjecthaving a cancer with aberrant H3-K27 methylation effective amount of oneor more compounds of Formula (I), (Ia), and/or (Ib), wherein thecompound(s) inhibits histone methyltransferase activity of EZH2, therebytreating the cancer. Examples of aberrant H3-K27 methylation may includea global increase in and/or altered distribution of H3-K27 di ortri-methylation within the cancer cell chromatin.

For example, the cancer is selected from the group consisting of cancersthat overexpress EZH2 or other PRC2 subunits, contain loss-of-functionmutations in H3-K27 demethylases such as UTX, or overexpress accessoryproteins such as PHF19/PCL3 capable of increasing and or mislocalizingEZH2 activity (see references in Sneeringer et al. Proc Natl Acad SciUSA 107(49):20980-5, 2010).

For example, the method comprises the step of administering to a subjecthaving a cancer overexpressing EZH2 a therapeutically effective amountof one or more compounds of Formula (I), (Ia), and/or (Ib), wherein thecompound(s) inhibits histone methyltransferase activity of EZH2, therebytreating the cancer.

For example, the method comprises the step of administering to a subjecthaving a cancer with a loss-of-function mutation in the H3-K27demethylase UTX a therapeutically effective amount of one or morecompounds of Formula (I), (Ia), and/or (Ib), wherein the compound(s)inhibits histone methyltransferase activity of EZH2, thereby treatingthe cancer

For example, the method comprises the step of administering to a subjecthaving a cancer overexpressing an accessory component(s) of the PRC2,such as PHF19/PCL3, a therapeutically effective amount of one or morecompounds of Formula (I), (Ia), and/or (Ib), wherein the compound(s)inhibits histone methyltransferase activity of EZH2, thereby treatingthe cancer

In still another aspect, this invention relates to a method ofmodulating the activity of the wild-type EZH2, the catalytic subunit ofthe PRC2 complex which catalyzes the mono- through tri-methylation oflysine 27 on histone H3 (H3-K27). For example, the present inventionrelates to a method of inhibiting the activity of EZH2 in a cell.

In yet another aspect, this invention features to a method of inhibitingin a subject conversion of H3-K27 to trimethylated H3-K27. The methodcomprises administering to a subject a therapeutically effective amountof one or more of the compound of Formula (I), (Ia), or (Ib) to inhibithistone methyltransferase activity of EZH2, thereby inhibitingconversion of H3-K27 to trimethylated H3-K27 in the subject.

For example, the method comprises the step of administering to a subjecthaving a cancer expressing a Y641 mutant of EZH2 a therapeuticallyeffective amount of one or more compounds of Formula (I), (Ia), and/or(Ib), wherein the compound(s) inhibits histone methyltransferaseactivity of EZH2, thereby treating the cancer.

For example, the cancer is selected from the group consisting offollicular lymphoma and diffuse large B-cell lymphoma (DLBCL) ofgerminal center B cell-like (GCB) subtype.

For example, the method comprises the step of administering to a subjecthaving a cancer expressing a Y641 mutant of EZH2 a therapeuticallyeffective amount of one or more compounds of Formula (I), (Ia), and/or(Ib), wherein the compound(s) selectively inhibits histonemethyltransferase activity of the Y641 mutant of EZH2, thereby treatingthe cancer.

For example, the method further comprises the steps of performing anassay to detect a Y641 mutant of EZH2 in a sample comprising cancercells from a subject having a cancer.

In still another aspect, this invention relates to a method ofmodulating the activity of the wild-type and mutant histonemethyltransferase EZH2, the catalytic subunit of the PRC2 complex whichcatalyzes the mono- through tri-methylation of lysine 27 on histone H3(H3-K27). For example, the present invention relates to a method ofinhibiting the activity of certain mutant forms of EZH2 in a cell. Themutant forms of EZH2 include a substitution of another amino acidresidue for tyrosine 641 (Y641, also Tyr641) of wild-type EZH2. Themethod includes contacting the cell with an effective amount of one ormore of the compound of Formula (I), (Ia), or (Ib).

In yet another aspect, this invention features to a method of inhibitingin a subject conversion of H3-K27 to trimethylated H3-K27. The methodcomprises administering to a subject expressing a Y641 mutant of EZH2 atherapeutically effective amount of one or more of the compound ofFormula (I), (Ia), or (Ib) to inhibit histone methyltransferase activityof EZH2, thereby inhibiting conversion of H3-K27 to trimethylated H3-K27in the subject. For example, the histone methyltransferase activityinhibited is that of the Y641 mutant of EZH2. For example, the compoundof this invention selectively inhibits histone methyltransferaseactivity of the Y641 mutant of EZH2. For example, the Y641 mutant ofEZH2 is selected from the group consisting of Y641C, Y641F, Y641H,Y641N, and Y641S.

The method of inhibiting in a subject conversion of H3-K27 totrimethylated H3-K27 may also comprise performing an assay to detect aY641 mutant of EZH2 in a sample from a subject before administering tothe subject expressing a Y641 mutant of EZH2 a therapeutically effectiveamount of one or more of the compound of Formula (I), (Ia), or (Ib). Forexample, performing the assay to detect the Y641 mutant of EZH2 includeswhole-genome resequencing or target region resequencing that detects anucleic acid encoding the Y641 mutant of EZH2. For example, performingthe assay to detect the Y641 mutant of EZH2 includes contacting thesample with an antibody that binds specifically to a polypeptide orfragment thereof characteristic of the Y641 mutant of EZH2. For example,performing the assay to detect the Y641 mutant of EZH2 includescontacting the sample under highly stringent conditions with a nucleicacid probe that hybridizes to a nucleic acid encoding a polypeptide orfragment thereof characteristic of the Y641 mutant of EZH2.

Further, the invention also relates to a method of identifying aninhibitor of a Y641 mutant of EZH2. The method comprises the steps ofcombining an isolated Y641 mutant of EZH2 with a histone substrate, amethyl group donor, and a test compound, wherein the histone substratecomprises a form of H3-K27 selected from the group consisting ofunmethylated H3-K27, monomethylated H3-K27, dimethylated H3-K27, and anycombination thereof; and performing an assay to detect methylation ofH3-K27 (e.g., formation of trimethylated H3-K27) in the histonesubstrate, thereby identifying the test compound as an inhibitor of theY641 mutant of EZH2 when methylation of H3-K27 (e.g., formation oftrimethylated H3-K27) in the presence of the test compound is less thanmethylation of H3-K27 (e.g., formation of trimethylated H3-K27) in theabsence of the test compound.

In one embodiment, performing the assay to detect methylation of H3-K27in the histone substrate comprises measuring incorporation of labeledmethyl groups.

In one embodiment, the labeled methyl groups are isotopically labeledmethyl groups.

In one embodiment, performing the assay to detect methylation of H3-K27in the histone substrate comprises contacting the histone substrate withan antibody that binds specifically to trimethylated H3-K27.

Also within the scope of the invention is a method of identifying aselective inhibitor of a Y641 mutant of EZH2. The method comprises thesteps of combining an isolated Y641 mutant of EZH2 with a histonesubstrate, a methyl group donor, and a test compound, wherein thehistone substrate comprises a form of H3-K27 selected from the groupconsisting of monomethylated H3-K27, dimethylated H3-K27, and acombination of monomethylated H3-K27 and dimethylated H3-K27, therebyforming a test mixture; combining an isolated wild-type EZH2 with ahistone substrate, a methyl group donor, and a test compound, whereinthe histone substrate comprises a form of H3-K27 selected from the groupconsisting of monomethylated H3-K27, dimethylated H3-K27, and acombination of monomethylated H3-K27 and dimethylated H3-K27, therebyforming a control mixture; performing an assay to detect trimethylationof the histone substrate in each of the test mixture and the controlmixture; calculating the ratio of (a) trimethylation with the Y641mutant of EZH2 and the test compound (M+) to (b) trimethylation with theY641 mutant of EZH2 without the test compound (M−); calculating theratio of (c) trimethylation with wild-type EZH2 and the test compound(WT+) to (d) trimethylation with wild-type EZH2 without the testcompound (WT−); comparing the ratio (a)/(b) with the ratio (c)/(d); andidentifying the test compound as a selective inhibitor of the Y641mutant of EZH2 when the ratio (a)/(b) is less than the ratio (c)/(d).

The present invention further provides a method of identifying a subjectas a candidate for treatment with one or more compounds of theinvention. The method comprises the steps of performing an assay todetect a Y641 mutant of EZH2 in a sample from a subject; and identifyinga subject expressing a Y641 mutant of EZH2 as a candidate for treatmentwith one or more compounds of the invention, wherein the compound(s)inhibits histone methyltransferase activity of EZH2.

Still another aspect of the invention is a method of inhibitingconversion of H3-K27 to trimethylated H3-K27. The method comprises thestep of contacting a Y641 mutant of EZH2 with a histone substratecomprising H3-K27 and an effective amount of a compound of the presentinvention, wherein the compound inhibits histone methyltransferaseactivity of EZH2, thereby inhibiting conversion of H3-K27 totrimethylated H3-K27.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. In the specification, thesingular forms also include the plural unless the context clearlydictates otherwise. Although methods and materials similar or equivalentto those described herein can be used in the practice or testing of thepresent invention, suitable methods and materials are described below.All publications, patent applications, patents and other referencesmentioned herein are incorporated by reference. The references citedherein are not admitted to be prior art to the claimed invention. In thecase of conflict, the present specification, including definitions, willcontrol. In addition, the materials, methods and examples areillustrative only and are not intended to be limiting.

Other features and advantages of the invention will be apparent from thefollowing detailed description and claims.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel substituted 6,5-fused bicyclicheteroaryl compounds, synthetic methods for making the compounds,pharmaceutical compositions containing them and various uses of thecompounds.

1. Substituted 6,5-fused Bicyclic Heteroaryl Compounds

The present invention provides the compounds of Formula (I):

or a pharmaceutically acceptable salt or ester thereof. In this formula:

-   -   X₁ is NR₇ or CR₇;    -   X₂ is N, NR₈, CR₈, O, or S;    -   X₃ is NR₈, CR₈, O, or S;    -   X₄ is C or N;    -   Y₁ is N or CH;    -   Y₂ is N or CR₆;    -   Y₃ is N, or CR₁₁, and at least one of X₁, X₂, X₃, X₄, Y₁, Y₂,        and Y₃ is N or NR₇;    -   each of R₁, R₅, R₉, and R₁₀, independently, is H or C₁-C₆ alkyl        optionally substituted with one or more substituents selected        from the group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆        alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, and C₃-C₈ cycloalkyl;    -   each of R₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₁ is H, halo, hydroxyl,        COOH, cyano, or R_(S1), in which R_(S1) is C₁-C₃ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl, C(O)O—C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and R_(S1) is optionally substituted with one or        more substituents selected from the group consisting of halo,        hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl,        amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5        to 6-membered heteroaryl;    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which each of        R_(a) and R_(b), independently is H or R_(S3) and each of R_(S2)        and R_(S3), independently is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl; or R_(a) and        R_(b), together with the N atom to which they are attached, form        a 4 to 7-membered heterocycloalkyl ring having 0 or 1 additional        heteroatoms to the N atom; and each of R_(S2), R_(S3), and the 4        to 7-membered heterocycloalkyl ring containing R_(a) and R_(b),        is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is        a bond or C₁-C₃ alkyl linker each optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano,        —OR_(c), —NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c),        —C(O)OR_(c), —C(O)NR_(c)R_(d), —NR_(d)C(O)R_(c),        —NR_(d)C(O)OR_(c), —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in        which each of R_(c) R_(d), and R_(d′), independently is H or        R_(S5), A⁻ is a pharmaceutically acceptable anion, each of        R_(S4) and R_(S5), independently, is C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, or 5        to 6-membered heteroaryl, or R_(c) and R_(d), together with the        N atom to which they are attached, form a 4 to 7-membered        heterocycloalkyl ring having 0 or 1 additional heteroatoms to        the N atom, and each of R_(S4), R_(S5), and the 4 to 7-membered        heterocycloalkyl ring containing R_(c) and R_(d), is optionally        substituted with one or more -Q₃-T₃, wherein Q₃ is a bond or        C₁-C₃ alkyl linker each optionally substituted with halo, cyano,        hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from the group        consisting of H, halo, cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, 5 to 6-membered        heteroaryl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and        —C(O)NR_(e)R_(f), each of R_(e) and R_(f) independently being H        or C₁-C₆ alkyl; or -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two        neighboring -Q₂-T₂, together with the atoms to which they are        attached form a 5- or 6-membered ring optionally containing 1-4        heteroatoms selected from N, O and S and optionally substituted        with one or more substituents selected from the group consisting        of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl,        and 5 to 6-membered heteroaryl; provided that -Q₂-T₂ is not H;    -   each R₇ independently is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄        alkyl linker, or C₂-C₄ alkenyl linker, each linker optionally        substituted with halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄        is H, halo, cyano, NR_(g)R_(h), —OR_(g), —C(O)R_(g),        —C(O)OR_(g), —C(O)NR_(g)R_(h), —C(O)NR_(g)OR_(h),        —NR_(g)C(O)R_(h), —S(O)₂R_(g), or R_(S6), in which each of R_(g)        and R_(h), independently is H or R_(S7), each of R_(S6) and        R_(S7), independently is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and each of R_(S6) and R_(S7) is optionally        substituted with one or more -Q₅-T₅, wherein Q₅ is a bond, C(O),        or C₁-C₃ alkyl linker, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl and T₅ is        optionally substituted with one or more substituents selected        from the group consisting of halo, C₁-C₆ alkyl, hydroxyl, cyano,        C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, and 5 to 6-membered heteroaryl except when T₅        is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; provided that        -Q₄-T₄ is not H; and    -   each of R₈ and R₁₁, independently, is H, halo, hydroxyl, COOH,        cyano, R_(S8), OR_(S8), or COOR_(S8), in which R_(S8) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S8) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino;    -   provided that the compound is not    -   N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,5-dimethylthiophen-3-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-(2,3-dihydro-1,4-benzodioxin-6-yl)-N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1,3-dimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1,3,6-trimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-6-methyl-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,        or    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide.

For example, at least one of Y₁, Y₃, and X₄ is N and when X₄ is C, Y₁ isN, Y₂ is CR₆, and Y₃ is CR₁₁ then X₂ is CR₈.

For example,

For example, X₄ is C.

For example, X₂ is N or CH.

For example, X₃ is CR₈.

For example, Y₃ is CR₁₁.

For example, R₆ is phenyl substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing 1-3 additionalheteroatoms selected from N, O, and S and optionally substituted withone or more -Q₂-T₂, provided that the heteroaryl is not thiophenyl.

For example, R₆ is pyridinyl, pyrazolyl, pyrimidinyl, or furyl, each ofwhich is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, R₆ is C₁-C₃ alkyl substituted with one or more -Q₂-T₂.

For example, R₆ is C₂-C₆ alkenyl or C₄-C₆ cycloalkyl each optionallysubstituted with one or more -Q₂-T₂.

For example, R₆ is C(O)H.

For example, R₆ is OR_(a) or —C(O)R_(a).

For example, R_(a) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is —NR_(a)R_(b), —C(O)R_(a), —C(O)OR_(a),—C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a), —S(O)₂R_(a), or —S(O)₂NR_(a)R_(b).

For example, each of R_(a) and R_(b), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₂-T₂.

For example, one of R_(a) and R_(b) is H.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more

-Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₂-T₂.

For example, R₆ is piperidinyl, 2,2,6,6-tetramethyl-piperidinyl,1,2,3,6-tetrahydropyridinyl,2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, orpyrrolidinyl, each of which is optionally substituted with one or more-Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl optionallysubstituted with one or more -Q₂-T₂, and -Q₂-T₂ is oxo or Q₂ is a bondand T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c),

—S(O)₂R_(c), C₁-C₆ alkyl, or 4 to 7-membered heterocycloalkyl, each ofwhich is optionally substituted with one or more -Q₃-T₃ when R_(c) orR_(d) is not H.

For example, -Q₂-T₂ is oxo.

For example, Q₂ is a bond.

For example, Q₂ is an unsubstituted C₁-C₃ alkyl linker.

For example, T₂ is C₁-C₆ alkyl or C₆-C₁₀ aryl, each optionallysubstituted with one or more -Q₃-T₃.

For example, T₂ is an unsubstituted substituted straight chain C₁-C₆ orbranched C₃-C₆ alkyl, including but not limited to, methyl, ethyl,n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl andn-hexyl.

For example, T₂ is phenyl.

For example, T₂ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, T₂ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₃-T₃.

For example, T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), or—S(O)₂R_(c).

For example, T₂ is —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)NR_(c)R_(d),—NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c), or —S(O)₂NR_(c)R_(d).

For example, Q₂ is a bond or methyl linker and T₂ is H, halo, —OR_(c),—NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, or —S(O)₂NR_(c)R_(d).

For example, R_(c) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₃-T₃.

For example, each of R_(c) and R_(d), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₃-T₃.

For example, R_(c) is H.

For example, R_(d) is H.

For example, R_(c) and R_(d), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more

-Q₃-T₃.

For example, Q₂ is a bond and T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c),—C(O)OR_(c), —S(O)₂R_(c), C₁-C₆ alkyl, or 4 to 7-memberedheterocycloalkyl, each of which is optionally substituted with one ormore -Q₃-T₃ when R_(c) or R_(d) is not H.

For example, -Q₃-T₃ is oxo.

For example, T₂ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₃-T₃ are oxo.

For example, Q₃ is a bond or unsubstituted or substituted C₁-C₃ alkyllinker.

For example, T₃ is H, halo, 4 to 7-membered heterocycloalkyl, C₁-C₃alkyl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), or —C(O)NR_(e)R_(f).

For example, one of R_(d) and R_(e) is H.

For example, Q₃ is a bond or C₁-C₃ alkyl linker and T₃ is selected fromthe group consisting of C₁-C₃ alkyl, halo, OR_(e), —S(O)₂R_(e),—NR_(e)R_(f), and —C(O)NR_(e)R_(f).

For example, R_(e) is H.

For example, R_(f) is H.

For example, R₇ is C₁-C₆ alkyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₃-C₈ cycloalkyl optionally substituted with one ormore -Q₅-T₅.

For example, R₇ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more

-Q₅-T₅.

For example, R₇ is cyclopentyl.

For example, R₇ is isopropyl.

For example, R₇ is 5 to 6-membered heterocycloalkyl optionallysubstituted with one or more -Q₅-T₅.

For example, R₇ is piperidinyl optionally substituted with one -Q₅-T₅.

For example, -Q₅-T₅ is oxo.

For example, T₄ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₅-T₅ are oxo.

For example, T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₅ is CO and T₅ is C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₅ is C₁-C₆ alkyl optionally substituted with halo,hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

For example, R₁₁ is H.

For example, each of R₂ and R₄, independently, is H or C₁-C₆ alkyloptionally substituted with amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₆-C₁₀ aryl.

For example, each of R₂ and R₄, independently is C₁-C₃ alkyl optionallysubstituted with C₁-C₆ alkoxyl.

For example, each of R₂ and R₄ is methyl.

For example, R₁ is H.

For example, R₅ is H, methyl, or ethyl.

For example, R₃ is H.

For example, each of R₅, R₉, and R₁₀ is H.

For example, A⁻ is Br⁻.

For example,

is selected from the group consisting of indolyl, isoindolyl,indolizinyl, benzofuryl, isobenzofuryl, benzo[b]thienyl, benzoxazolyl,benzthiazolyl, benzimidazolyl, benzotriazolyl, benzoxadiazolyl,benzothiadiazolyl, purinyl, indazolyl, pyrrolopyridinyl,imidazopyridinyl, pyrazolopyridinyl, pyrrolopyrazinyl, imidazopyrazinyl,pyrazolopyrazinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl,pyrrolopyridazinyl, imidazopyridazinyl, pyrazolopyridazinyl,furopyridinyl, thienopyridinyl, furopyrazinyl, thienopyrazinyl,oxazolopyridinyl, isoxazolopyridinyl, thiazolopyridinyl,isothiazolopyridinyl, oxadiazolopyridinyl, thiadiazolopyridinyl,triazolopyridinyl, oxazolopyrazinyl, isoxazolopyrazinyl,thiazolopyrazinyl, isothiazolopyrazinyl, oxadiazolopyrazinyl,thiadiazolopyrazinyl, triazolopyrazinyl, furopyrimidinyl,thienopyrimidinyl, furopyridazinyl, thienopyridazinyl,oxazolopyrimidinyl, isoxazolopyrimidinyl, thiazolopyrimidinyl,isothiazolopyrimidinyl, oxadiazolopyrimidinyl, thiadiazolopyrimidinyl,triazolopyrimidinyl, oxazolopyridazinyl, isoxazolopyridazinyl,thiazolopyridazinyl, isothiazolopyridazinyl, oxadiazolopyridazinyl,thiadiazolopyridazinyl, triazolopyridazinyl, and imidazotriazinyl.

For example,

is selected from the group consisting of

For example,

is selected from the group consisting of

For example,

is selected from the group consisting of

In another aspect, the present invention features substituted 6,5-fusedbicyclic heteroaryl compound of Formula (I):

or a pharmaceutically acceptable salt or ester thereof, in which

-   -   X₁ is NR₇ or CR₇;    -   X₂ is N, NR₈, CR₈, O, or S;    -   X₃ is NR₈, CR₈, O, or S;    -   X₄ is C or N;    -   Y₁ is N or CH;    -   Y₂ is N or CR₆;    -   Y₃ is N, or CR₁₁, and at least one of X₁, X₂, X₃, X₄, Y₁, Y₂,        and Y₃ is N or NR₇;    -   each of R₁, R₅, R₉, and R₁₀, independently, is H or C₁-C₆ alkyl        optionally substituted with one or more substituents selected        from the group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆        alkyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino,        di-C₁-C₆ alkylamino, and C₃-C₈ cycloalkyl;    -   each of R₂, R₃, and R₄, independently, is -Q₁-T₁, in which Q₁ is        a bond or C₁-C₃ alkyl linker optionally substituted with halo,        cyano, hydroxyl or C₁-C₆ alkoxy, and T₁ is H, halo, hydroxyl,        COOH, cyano, or R_(S1), in which R_(S1) is C₁-C₃ alkyl, C₂-C₆        alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkoxyl, C(O)O—C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and R_(S1) is optionally substituted with one or        more substituents selected from the group consisting of halo,        hydroxyl, oxo, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl,        amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5        to 6-membered heteroaryl;    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which R_(S2) is        C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl        containing at least one N or O atom, each of R_(a) and R_(b),        independently is H or R_(S3), and R_(S3), independently is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl; or R_(a) and R_(b), together with the N atom to        which they are attached, form a 4 to 7-membered heterocycloalkyl        ring having 0 or 1 additional heteroatoms to the N atom; and        each of R_(S2), R_(S3), and the 4 to 7-membered heterocycloalkyl        ring containing R_(a) and R_(b), is optionally substituted with        one or more -Q₂-T₂, wherein Q₂ is a bond or C₁-C₃ alkyl linker        each optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₂ is H, halo, cyano, —NR_(c)R_(d),        —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c), —C(O)OR_(c),        —C(O)NR_(c)R_(d), —NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c),        —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in which each of        R_(c) R_(d), and R_(d′), independently is H or R_(S5), A⁻ is a        pharmaceutically acceptable anion, each of R_(S4) and R_(S5),        independently, is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl,        or R_(c) and R_(d), together with the N atom to which they are        attached, form a 4 to 7-membered heterocycloalkyl ring having 0        or 1 additional heteroatoms to the N atom, and each of R_(S4),        R_(S5), and the 4 to 7-membered heterocycloalkyl ring containing        R_(c) and R_(d), is optionally substituted with one or more        -Q₃-T₃, wherein Q₃ is a bond or C₁-C₃ alkyl linker each        optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₃ is selected from the group consisting of halo,        cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        7-membered heterocycloalkyl, 5 to 6-membered heteroaryl, OR_(e),        COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and —C(O)NR_(e)R_(f), each        of R_(e) and R_(f) independently being H or C₁-C₆ alkyl; or        -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two neighboring -Q₂-T₂,        together with the atoms to which they are attached form a 5- or        6-membered ring optionally containing 1-4 heteroatoms selected        from N, O and S and optionally substituted with one or more        substituents selected from the group consisting of halo,        hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,        mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5 to        6-membered heteroaryl; provided that -Q₂-T₂ is not H;    -   each R₇ independently is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄        alkyl linker, or C₂-C₄ alkenyl linker, each linker optionally        substituted with halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄        is H, halo, cyano, NR_(g)R_(h), —OR_(g), —C(O)R_(g),        —C(O)OR_(g), —C(O)NR_(g)R_(h), —C(O)NR_(g)OR_(h),        —NR_(g)C(O)R_(h), —S(O)₂R_(g), or R_(S6), in which each of R_(g)        and R_(h), independently is H or R_(S7), each of R_(S6) and        R_(S7), independently is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and each of R_(S6) and R_(S7) is optionally        substituted with one or more -Q₅-T₅, wherein Q₅ is a bond, C(O),        or C₁-C₃ alkyl linker, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl and T₅ is        optionally substituted with one or more substituents selected        from the group consisting of halo, C₁-C₆ alkyl, hydroxyl, cyano,        C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, and 5 to 6-membered heteroaryl except when T₅        is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; provided that        -Q₄-T₄ is not H; and    -   each of R₈ and R₁₁, independently, is H, halo, hydroxyl, COOH,        cyano, R_(S8), OR_(S8), or COOR_(S8), in which R_(S8) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S8) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino.

For example, X₄ is C.

For example, X₂ is N or CH.

For example, X₃ is CR₈.

For example, Y₃ is CR₁₁.

For example, R₆ is phenyl substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing at least one Nor O atom and is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is pyridinyl, pyrazolyl, pyrimidinyl, or furyl, each ofwhich is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, R₆ is C₂-C₆ alkenyl optionally substituted with one or more-Q₂-T₂.

For example, R₆ is C(O)H.

For example, R₆ is OR_(a) or —C(O)R_(a).

For example, R_(a) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is —NR_(a)R_(b), —C(O)R_(a), —C(O)OR_(a),—C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a), —S(O)₂R_(a), or —S(O)₂NR_(a)R_(b).

For example, each of R_(a) and R_(b), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₂-T₂.

For example, one of R_(a) and R_(b) is H.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₂-T₂.

For example, R₆ is piperidinyl, 2,2,6,6-tetramethyl-piperidinyl,1,2,3,6-tetrahydropyridinyl,2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, orpyrrolidinyl, each of which is optionally substituted with one or more-Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl optionallysubstituted with one or more -Q₂-T₂, and -Q₂-T₂ is oxo or Q₂ is a bondand T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), —S(O)₂R_(c),C₁-C₆ alkyl, or 4 to 7-membered heterocycloalkyl, each of which isoptionally substituted with one or more -Q₃-T₃ when R_(c) or R_(d) isnot H.

For example, -Q₂-T₂ is oxo.

For example, Q₂ is a bond.

For example, Q₂ is an unsubstituted C₁-C₃ alkyl linker.

For example, T₂ is C₁-C₆ alkyl or C₆-C₁₀ aryl, each optionallysubstituted with one or more -Q₃-T₃.

For example, T₂ is an unsubstituted substituted straight chain C₁-C₆ orbranched C₃-C₆ alkyl, including but not limited to, methyl, ethyl,n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl andn-hexyl.

For example, T₂ is phenyl.

For example, T₂ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, T₂ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₃-T₃.

For example, T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), or—S(O)₂R_(c).

For example, T₂ is —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)NR_(c)R_(d),—NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c), or —S(O)₂NR_(c)R_(d).

For example, Q₂ is a bond or methyl linker and T₂ is H, halo, —OR_(c),—NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, or —S(O)₂NR_(c)R_(d).

For example, R_(c) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₃-T₃.

For example, each of R_(c) and R_(d), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₃-T₃.

For example, R_(c) is H.

For example, R_(d) is H.

For example, R_(c) and R_(d), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more -Q₃-T₃.

For example, Q₂ is a bond and T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c),—C(O)OR_(e), —S(O)₂R_(c), C₁-C₆ alkyl, or 4 to 7-memberedheterocycloalkyl, each of which is optionally substituted with one ormore -Q₃-T₃ when R_(c) or R_(d) is not H.

For example, -Q₃-T₃ is oxo.

For example, T₂ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₃-T₃ are oxo.

For example, Q₃ is a bond or unsubstituted or substituted C₁-C₃ alkyllinker.

For example, T₃ is H, halo, 4 to 7-membered heterocycloalkyl, C₁-C₃alkyl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), or —C(O)NR_(e)R_(f).

For example, one of R_(d) and R_(e) is H.

For example, Q₃ is a bond or C₁-C₃ alkyl linker and T₃ is selected fromthe group consisting of C₁-C₃ alkyl, halo, OR_(e), —S(O)₂R_(e),—NR_(e)R_(f), and —C(O)NR_(e)R_(f).

For example, R_(e) is H.

For example, R_(f) is H.

For example, R₇ is C₁-C₆ alkyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₃-C₈ cycloalkyl optionally substituted with one ormore -Q₅-T₅.

For example, R₇ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₅-T₅.

For example, R₇ is cyclopentyl.

For example, R₇ is isopropyl.

For example, R₇ is 5 to 6-membered heterocycloalkyl optionallysubstituted with one or more -Q₅-T₅.

For example, R₇ is piperidinyl optionally substituted with one -Q₅-T₅.

For example, -Q₅-T₅ is oxo.

For example, T₄ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₅-T₅ are oxo.

For example, T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₅ is CO and T₅ is C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₅ is C₁-C₆ alkyl optionally substituted with halo,hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

For example, R₁₁ is H.

For example, each of R₂ and R₄, independently, is H or C₁-C₆ alkyloptionally substituted with amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₆-C₁₀ aryl.

For example, each of R₂ and R₄, independently is C₁-C₃ alkyl optionallysubstituted with C₁-C₆ alkoxyl.

For example, each of R₂ and R₄ is methyl.

For example, R₁ is H.

For example, R₅ is H, methyl, or ethyl.

For example, R₃ is H.

For example, each of R₅, R₉, and R₁₀ is H.

For example, A⁻ is Br⁻.

For example,

is selected from the group consisting of indolyl, isoindolyl,indolizinyl, benzofuryl, isobenzofuryl, benzo[b]thienyl, benzoxazolyl,benzthiazolyl, benzimidazolyl, benzotriazolyl, benzoxadiazolyl,benzothiadiazolyl, purinyl, indazolyl, pyrrolopyridinyl,imidazopyridinyl, pyrazolopyridinyl, pyrrolopyrazinyl, imidazopyrazinyl,pyrazolopyrazinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl,pyrrolopyridazinyl, imidazopyridazinyl, pyrazolopyridazinyl,furopyridinyl, thienopyridinyl, furopyrazinyl, thienopyrazinyl,oxazolopyridinyl, isoxazolopyridinyl, thiazolopyridinyl,isothiazolopyridinyl, oxadiazolopyridinyl, thiadiazolopyridinyl,triazolopyridinyl, oxazolopyrazinyl, isoxazolopyrazinyl,thiazolopyrazinyl, isothiazolopyrazinyl, oxadiazolopyrazinyl,thiadiazolopyrazinyl, triazolopyrazinyl, furopyrimidinyl,thienopyrimidinyl, furopyridazinyl, thienopyridazinyl,oxazolopyrimidinyl, isoxazolopyrimidinyl, thiazolopyrimidinyl,isothiazolopyrimidinyl, oxadiazolopyrimidinyl, thiadiazolopyrimidinyl,triazolopyrimidinyl, oxazolopyridazinyl, isoxazolopyridazinyl,thiazolopyridazinyl, isothiazolopyridazinyl, oxadiazolopyridazinyl,thiadiazolopyridazinyl, triazolopyridazinyl, and imidazotriazinyl.

For example,

is selected from the group consisting of

For example,

is selected from the group consisting of

For example,

is selected from the group consisting of

The present invention provides the compounds of Formula (Ia)

or a pharmaceutically acceptable salt or ester thereof, wherein:

-   -   X₁ is NR₇ or CR₇;    -   X₂ is N, NR₈, CR₈, O, or S;    -   X₃ is NR₈, CR₈, O, or S;    -   X₄ is C or N;    -   Y₁ is N or CH;    -   Y₂ is N or CR₆;    -   Y₃ is N, or CR₁₁, and at least one of X₁, X₂, X₃, X₄, Y₁, Y₂,        and Y₃ is N or NR₇;    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which each of        R_(a) and R_(b), independently is H or R_(S3) and each of R_(S2)        and R_(S3), independently is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl; or R_(a) and        R_(b), together with the N atom to which they are attached, form        a 4 to 7-membered heterocycloalkyl ring having 0 or 1 additional        heteroatoms to the N atom; and each of R_(S2), R_(S3), and the 4        to 7-membered heterocycloalkyl ring containing R_(a) and R_(b),        is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is        a bond or C₁-C₃ alkyl linker each optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano,        —OR_(c), —NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c),        —C(O)OR_(c), —C(O)NR_(c)R_(d), —NR_(d)C(O)R_(c),        —NR_(d)C(O)OR_(c), —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in        which each of R_(c) R_(d), and R_(d′), independently is H or        R_(S5), A⁻ is a pharmaceutically acceptable anion, each of        R_(S4) and R_(S5), independently, is C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, or 5        to 6-membered heteroaryl, or R_(c) and R_(d), together with the        N atom to which they are attached, form a 4 to 7-membered        heterocycloalkyl ring having 0 or 1 additional heteroatoms to        the N atom, and each of R_(S4), R_(S5), and the 4 to 7-membered        heterocycloalkyl ring containing R_(c) and R_(d), is optionally        substituted with one or more -Q₃-T₃, wherein Q₃ is a bond or        C₁-C₃ alkyl linker each optionally substituted with halo, cyano,        hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from the group        consisting of H, halo, cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, 5 to 6-membered        heteroaryl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and        —C(O)NR_(e)R_(f), each of R_(e) and R_(f) independently being H        or C₁-C₆ alkyl; or -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two        neighboring -Q₂-T₂, together with the atoms to which they are        attached form a 5- or 6-membered ring optionally containing 1-4        heteroatoms selected from N, O and S and optionally substituted        with one or more substituents selected from the group consisting        of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl,        and 5 to 6-membered heteroaryl; provided that -Q₂-T₂ is not H;        or    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which R_(S2) is        C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl        containing at least one N or O atom, each of R_(a) and R_(b),        independently is H or R_(S3), and R_(S3), independently is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl; or R_(a) and R_(b), together with the N atom to        which they are attached, form a 4 to 7-membered heterocycloalkyl        ring having 0 or 1 additional heteroatoms to the N atom; and        each of R_(S2), R_(S3), and the 4 to 7-membered heterocycloalkyl        ring containing R_(a) and R_(b), is optionally substituted with        one or more -Q₂-T₂, wherein Q₂ is a bond or C₁-C₃ alkyl linker        each optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₂ is H, halo, cyano, —NR_(c)R_(d),        —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c), —C(O)OR_(c),        —C(O)NR_(c)R_(d), —NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c),        —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in which each of        R_(c) R_(d), and R_(d′), independently is H or R_(S5), A⁻ is a        pharmaceutically acceptable anion, each of R_(S4) and R_(S5),        independently, is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl,        or R_(c) and R_(d), together with the N atom to which they are        attached, form a 4 to 7-membered heterocycloalkyl ring having 0        or 1 additional heteroatoms to the N atom, and each of R_(S4),        R_(S5), and the 4 to 7-membered heterocycloalkyl ring containing        R_(c) and R_(d), is optionally substituted with one or more        -Q₃-T₃, wherein Q₃ is a bond or C₁-C₃ alkyl linker each        optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₃ is selected from the group consisting of halo,        cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        7-membered heterocycloalkyl, 5 to 6-membered heteroaryl, OR_(e),        COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and —C(O)NR_(e)R_(f), each        of R_(e) and R_(f) independently being H or C₁-C₆ alkyl; or        -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two neighboring -Q₂-T₂,        together with the atoms to which they are attached form a 5- or        6-membered ring optionally containing 1-4 heteroatoms selected        from N, O and S and optionally substituted with one or more        substituents selected from the group consisting of halo,        hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,        mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5 to        6-membered heteroaryl; provided that -Q₂-T₂ is not H;    -   each R₇ independently is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄        alkyl linker, or C₂-C₄ alkenyl linker, each linker optionally        substituted with halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄        is H, halo, cyano, NR_(g)R_(h), —OR_(g), —C(O)R_(g),        —C(O)OR_(g), —C(O)NR_(g)R_(h), —C(O)NR_(g)OR_(h),        —NR_(g)C(O)R_(h), —S(O)₂R_(g), or R_(S6), in which each of R_(g)        and R_(h), independently is H or R_(S7), each of R_(S6) and        R_(S7), independently is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and each of R_(S6) and R_(S7) is optionally        substituted with one or more -Q₅-T₅, wherein Q₅ is a bond, C(O),        or C₁-C₃ alkyl linker, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl and T₅ is        optionally substituted with one or more substituents selected        from the group consisting of halo, C₁-C₆ alkyl, hydroxyl, cyano,        C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, and 5 to 6-membered heteroaryl except when T₅        is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; provided that        -Q₄-T₄ is not H; and    -   each of R₈ and R₁₁, independently, is H, halo, hydroxyl, COOH,        cyano, R_(S8), OR_(S8), or COOR_(S8), in which R_(S8) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S8) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino;        provided that the compound is not    -   N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,5-dimethylthiophen-3-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-(2,3-dihydro-1,4-benzodioxin-6-yl)-N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1,3-dimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1,3,6-trimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-6-methyl-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,        or    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide.

For example, X₄ is C.

For example, X₂ is N or CH.

For example, X₃ is CR₈.

For example, Y₃ is CR₁₁.

For example, R₆ is phenyl substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing at least one Nor O atom and is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing 1-3 additionalheteroatoms selected from N, O, and S and optionally substituted withone or more -Q₂-T₂, provided that the heteroaryl is not thiophenyl.

For example, R₆ is pyridinyl, pyrazolyl, pyrimidinyl, or furyl, each ofwhich is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, R₆ is C₁-C₃ alkyl substituted with one or more -Q₂-T₂.

For example, R₆ is C₄-C₆ cycloalkyl optionally substituted with one ormore -Q₂-T₂.

For example, R₆ is C₂-C₆ alkenyl optionally substituted with one or more-Q₂-T₂.

For example, R₆ is C(O)H.

For example, R₆ is OR_(a) or —C(O)R_(a).

For example, R_(a) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is —NR_(a)R_(b), —C(O)R_(a), —C(O)OR_(a),—C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a), —S(O)₂R_(a), or —S(O)₂NR_(a)R_(b).

For example, each of R_(a) and R_(b), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₂-T₂.

For example, one of R_(a) and R_(b) is H.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₂-T₂.

For example, R₆ is piperidinyl, 2,2,6,6-tetramethyl-piperidinyl,1,2,3,6-tetrahydropyridinyl,2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, orpyrrolidinyl, each of which is optionally substituted with one or more-Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl optionallysubstituted with one or more -Q₂-T₂, and -Q₂-T₂ is oxo or Q₂ is a bondand T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), —S(O)₂R_(c),C₁-C₆ alkyl, or 4 to 7-membered heterocycloalkyl, each of which isoptionally substituted with one or more -Q₃-T₃ when R_(c) or R_(d) isnot H.

For example, -Q₂-T₂ is oxo.

For example, Q₂ is a bond.

For example, Q₂ is an unsubstituted C₁-C₃ alkyl linker.

For example, T₂ is C₁-C₆ alkyl or C₆-C₁₀ aryl, each optionallysubstituted with one or more -Q₃-T₃.

For example, T₂ is an unsubstituted substituted straight chain C₁-C₆ orbranched C₃-C₆ alkyl, including but not limited to, methyl, ethyl,n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl andn-hexyl.

For example, T₂ is phenyl.

For example, T₂ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, T₂ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₃-T₃.

For example, T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), or—S(O)₂R_(c).

For example, T₂ is —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)NR_(c)R_(d),—NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c), or —S(O)₂NR_(c)R_(d).

For example, Q₂ is a bond or methyl linker and T₂ is H, halo, —OR_(c),—NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, or —S(O)₂NR_(c)R_(d).

For example, R_(c) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₃-T₃.

For example, each of R_(c) and R_(d), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₃-T₃.

For example, R_(c) is H.

For example, R_(d) is H.

For example, R_(c) and R_(d), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more -Q₃-T₃.

For example, Q₂ is a bond and T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c),—C(O)OR_(c), —S(O)₂R_(c), C₁-C₆ alkyl, or 4 to 7-memberedheterocycloalkyl, each of which is optionally substituted with one ormore -Q₃-T₃ when R_(c) or R_(d) is not H.

For example, -Q₃-T₃ is oxo.

For example, T₂ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₃-T₃ are oxo.

For example, Q₃ is a bond or unsubstituted or substituted C₁-C₃ alkyllinker.

For example, T₃ is H, halo, 4 to 7-membered heterocycloalkyl, C₁-C₃alkyl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), or —C(O)NR_(e)R_(f).

For example, one of R_(d) and R_(e) is H.

For example, Q₃ is a bond or C₁-C₃ alkyl linker and T₃ is selected fromthe group consisting of C₁-C₃ alkyl, halo, OR_(e), —S(O)₂R_(e),—NR_(e)R_(f), and —C(O)NR_(e)R_(f).

For example, R_(e) is H.

For example, R_(f) is H.

For example, R₇ is C₁-C₆ alkyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₃-C₈ cycloalkyl optionally substituted with one ormore -Q₅-T₅.

For example, R₇ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₅-T₅.

For example, R₇ is cyclopentyl.

For example, R₇ is isopropyl.

For example, R₇ is 5 to 6-membered heterocycloalkyl optionallysubstituted with one or more -Q₅-T₅.

For example, R₇ is piperidinyl optionally substituted with one -Q₅-T₅.

For example, -Q₅-T₅ is oxo.

For example, T₄ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₅-T₅ are oxo.

For example, T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₅ is CO and T₅ is C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₅ is C₁-C₆ alkyl optionally substituted with halo,hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

For example, R₁₁ is H.

For example, R₅ is H, methyl, or ethyl.

For example, A⁻ is Br⁻.

For example,

For example,

is selected from the group consisting of

For example,

is selected from the group consisting of

The present invention also provides the compounds of Formula (Ib)

or a pharmaceutically acceptable salt or ester thereof, wherein:

-   -   X₂ is N, NR₈, CR₈, O, or S;    -   X₃ is NR₈, CR₈, O, or S;    -   X₄ is C or N;    -   Y₁ is N or CH;    -   Y₂ is N or CR₆;    -   Y₃ is N, or CR₁₁, and at least one of X₁, X₂, X₃, X₄, Y₁, Y₂,        and Y₃ is N or NR₇;    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which each of        R_(a) and R_(b), independently is H or R_(S3) and each of R_(S2)        and R_(S3), independently is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆        alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl; or R_(a) and        R_(b), together with the N atom to which they are attached, form        a 4 to 7-membered heterocycloalkyl ring having 0 or 1 additional        heteroatoms to the N atom; and each of R_(S2), R_(S3), and the 4        to 7-membered heterocycloalkyl ring containing R_(a) and R_(b),        is optionally substituted with one or more -Q₂-T₂, wherein Q₂ is        a bond or C₁-C₃ alkyl linker each optionally substituted with        halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₂ is H, halo, cyano,        —OR_(c), —NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c),        —C(O)OR_(c), —C(O)NR_(c)R_(d), —NR_(d)C(O)R_(c),        —NR_(d)C(O)OR_(c), —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in        which each of R_(c) R_(d), and R_(d′), independently is H or        R_(S5), A⁻ is a pharmaceutically acceptable anion, each of        R_(S4) and R_(S5), independently, is C₁-C₆ alkyl, C₃-C₈        cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, or 5        to 6-membered heteroaryl, or R_(c) and R_(d), together with the        N atom to which they are attached, form a 4 to 7-membered        heterocycloalkyl ring having 0 or 1 additional heteroatoms to        the N atom, and each of R_(S4), R_(S5), and the 4 to 7-membered        heterocycloalkyl ring containing R_(c) and R_(d), is optionally        substituted with one or more -Q₃-T₃, wherein Q₃ is a bond or        C₁-C₃ alkyl linker each optionally substituted with halo, cyano,        hydroxyl or C₁-C₆ alkoxy, and T₃ is selected from the group        consisting of H, halo, cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, 5 to 6-membered        heteroaryl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and        —C(O)NR_(e)R_(f), each of R_(e) and R_(f) independently being H        or C₁-C₆ alkyl; or -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two        neighboring -Q₂-T₂, together with the atoms to which they are        attached form a 5- or 6-membered ring optionally containing 1-4        heteroatoms selected from N, O and S and optionally substituted        with one or more substituents selected from the group consisting        of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆        alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino,        C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl,        and 5 to 6-membered heteroaryl; provided that -Q₂-T₂ is not H;        or    -   each R₆ independently is H, halo, OR_(a), —NR_(a)R_(b),        —C(O)R_(a), —C(O)OR_(a), —C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a),        —S(O)₂R_(a), —S(O)₂NR_(a)R_(b), or R_(S2), in which R_(S2) is        C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl        containing at least one N or O atom, each of R_(a) and R_(b),        independently is H or R_(S3), and R_(S3), independently is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl; or R_(a) and R_(b), together with the N atom to        which they are attached, form a 4 to 7-membered heterocycloalkyl        ring having 0 or 1 additional heteroatoms to the N atom; and        each of R_(S2), R_(S3), and the 4 to 7-membered heterocycloalkyl        ring containing R_(a) and R_(b), is optionally substituted with        one or more -Q₂-T₂, wherein Q₂ is a bond or C₁-C₃ alkyl linker        each optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₂ is H, halo, cyano, —NR_(c)R_(d),        —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)R_(c), —C(O)OR_(c),        —C(O)NR_(C)R_(d), —NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c),        —S(O)₂R_(c), —S(O)₂NR_(c)R_(d), or R_(S4), in which each of        R_(c) R_(d), and R_(d′), independently is H or R_(S5), A⁻ is a        pharmaceutically acceptable anion, each of R_(S4) and R_(S5),        independently, is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4        to 7-membered heterocycloalkyl, or 5 to 6-membered heteroaryl,        or R_(c) and R_(d), together with the N atom to which they are        attached, form a 4 to 7-membered heterocycloalkyl ring having 0        or 1 additional heteroatoms to the N atom, and each of R_(S4),        R_(S8), and the 4 to 7-membered heterocycloalkyl ring containing        R_(c) and R_(d), is optionally substituted with one or more        -Q₃-T₃, wherein Q₃ is a bond or C₁-C₃ alkyl linker each        optionally substituted with halo, cyano, hydroxyl or C₁-C₆        alkoxy, and T₃ is selected from the group consisting of halo,        cyano, C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to        7-membered heterocycloalkyl, 5 to 6-membered heteroaryl, OR_(e),        COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), and —C(O)NR_(e)R_(f), each        of R_(e) and R_(f) independently being H or C₁-C₆ alkyl; or        -Q₃-T₃ is oxo; or -Q₂-T₂ is oxo; or any two neighboring -Q₂-T₂,        together with the atoms to which they are attached form a 5- or        6-membered ring optionally containing 1-4 heteroatoms selected        from N, O and S and optionally substituted with one or more        substituents selected from the group consisting of halo,        hydroxyl, COOH, C(O)O—C₁-C₆ alkyl, cyano, C₁-C₆ alkoxyl, amino,        mono-C₁-C₆ alkylamino, di-C₁-C₆ alkylamino, C₃-C₈ cycloalkyl,        C₆-C₁₀ aryl, 4 to 7-membered heterocycloalkyl, and 5 to        6-membered heteroaryl; provided that -Q₂-T₂ is not H;    -   each R₇ independently is -Q₄-T₄, in which Q₄ is a bond, C₁-C₄        alkyl linker, or C₂-C₄ alkenyl linker, each linker optionally        substituted with halo, cyano, hydroxyl or C₁-C₆ alkoxy, and T₄        is H, halo, cyano, NR_(g)R_(h), —OR_(g), —C(O)R_(g),        —C(O)OR_(g), —C(O)NR_(g)R_(h), —C(O)NR_(g)OR_(h),        —NR_(g)C(O)R_(h), —S(O)₂R_(g), or R_(S6), in which each of R_(g)        and R_(h), independently is H or R_(S7), each of R_(S6) and        R_(S7), independently is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, C₆-C₁₀        aryl, 4 to 7-membered heterocycloalkyl, or 5 to 6-membered        heteroaryl, and each of R_(S6) and R_(S7) is optionally        substituted with one or more -Q₅-T₅, wherein Q₅ is a bond, C(O),        or C₁-C₃ alkyl linker, and T₅ is H, halo, C₁-C₆ alkyl, hydroxyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, or 5 to 6-membered heteroaryl and T₅ is        optionally substituted with one or more substituents selected        from the group consisting of halo, C₁-C₆ alkyl, hydroxyl, cyano,        C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆        alkylamino, C₃-C₈ cycloalkyl, C₆-C₁₀ aryl, 4 to 7-membered        heterocycloalkyl, and 5 to 6-membered heteroaryl except when T₅        is H, halo, hydroxyl, or cyano; or -Q₅-T₅ is oxo; provided that        -Q₄-T₄ is not H; and    -   each of R₈ and R₁₁, independently, is H, halo, hydroxyl, COOH,        cyano, R_(S8), OR_(S8), or COOR_(S8), in which R_(S8) is C₁-C₆        alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, amino, mono-C₁-C₆        alkylamino, or di-C₁-C₆ alkylamino, and R_(S8) is optionally        substituted with one or more substituents selected from the        group consisting of halo, hydroxyl, COOH, C(O)O—C₁-C₆ alkyl,        cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, and di-C₁-C₆        alkylamino;        provided that the compound is not    -   N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,5-dimethylthiophen-3-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-(2,3-dihydro-1,4-benzodioxin-6-yl)-N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1,3-dimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-1,3,6-trimethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,    -   N-[(1,2-dihydro-4,6-dimethyl-2-oxo-3-pyridinyl)methyl]-6-methyl-1-(1-methylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide,        or    -   6-cyclopropyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide.

For example, X₄ is C.

For example, X₂ is N or CH.

For example, X₃ is CR₈.

For example, Y₃ is CR₁₁.

For example, R₆ is phenyl substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing at least one Nor O atom and is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is 5 to 6-membered heteroaryl containing 1-3 additionalheteroatoms selected from N, O, and S and optionally substituted withone or more -Q₂-T₂, provided that the heteroaryl is not thiophenyl.

For example, R₆ is pyridinyl, pyrazolyl, pyrimidinyl, or furyl, each ofwhich is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, R₆ is C₁-C₃ alkyl substituted with one or more -Q₂-T₂.

For example, R₆ is C₄-C₆ cycloalkyl optionally substituted with one ormore -Q₂-T₂.

For example, R₆ is C₂-C₆ alkenyl optionally substituted with one or more-Q₂-T₂.

For example, R₆ is C(O)H.

For example, R₆ is OR_(a) or —C(O)R_(a).

For example, R_(a) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is —NR_(a)R_(b), —C(O)R_(a), —C(O)OR_(a),—C(O)NR_(a)R_(b), —NR_(b)C(O)R_(a), —S(O)₂R_(a), or —S(O)₂NR_(a)R_(b).

For example, each of R_(a) and R_(b), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₂-T₂.

For example, one of R_(a) and R_(b) is H.

For example, R_(a) and R_(b), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more -Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₂-T₂.

For example, R₆ is piperidinyl, 2,2,6,6-tetramethyl-piperidinyl,1,2,3,6-tetrahydropyridinyl,2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl, piperazinyl,morpholinyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, orpyrrolidinyl, each of which is optionally substituted with one or more-Q₂-T₂.

For example, R₆ is 4 to 7-membered heterocycloalkyl optionallysubstituted with one or more -Q₂-T₂, and -Q₂-T₂ is oxo or Q₂ is a bondand T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), —S(O)₂R_(c),C₁-C₆ alkyl, or 4 to 7-membered heterocycloalkyl, each of which isoptionally substituted with one or more -Q₃-T₃ when R_(c) or R_(d) isnot H.

For example, -Q₂-T₂ is oxo.

For example, Q₂ is a bond.

For example, Q₂ is an unsubstituted C₁-C₃ alkyl linker.

For example, T₂ is C₁-C₆ alkyl or C₆-C₁₀ aryl, each optionallysubstituted with one or more -Q₃-T₃.

For example, T₂ is an unsubstituted substituted straight chain C₁-C₆ orbranched C₃-C₆ alkyl, including but not limited to, methyl, ethyl,n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl andn-hexyl.

For example, T₂ is phenyl.

For example, T₂ is halo (e.g., fluorine, chlorine, bromine, and iodine).

For example, T₂ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₃-T₃.

For example, T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c), —C(O)OR_(c), or—S(O)₂R_(c).

For example, T₂ is —(NR_(c)R_(d)R_(d′))⁺A⁻, —C(O)NR_(c)R_(d),—NR_(d)C(O)R_(c), —NR_(d)C(O)OR_(c), or —S(O)₂NR_(c)R_(d).

For example, Q₂ is a bond or methyl linker and T₂ is H, halo, —OR_(c),—NR_(c)R_(d), —(NR_(c)R_(d)R_(d′))⁺A⁻, or —S(O)₂NR_(c)R_(d).

For example, R_(c) is C₁-C₆ alkyl or 4 to 7-membered heterocycloalkyl(e.g., azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl,pyrazolidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl,tetrahyrofuranyl, piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike), which is optionally substituted with one or more -Q₃-T₃.

For example, each of R_(c) and R_(d), independently is H or C₁-C₆ alkyloptionally substituted with one or more -Q₃-T₃.

For example, R_(c) is H.

For example, R_(d) is H.

For example, R_(c) and R_(d), together with the N atom to which they areattached, form a 4 to 7-membered heterocycloalkyl ring having 0 or 1additional heteroatoms to the N atom (e.g., azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, and morpholinyl, and the like)and the ring is optionally substituted with one or more -Q₃-T₃.

For example, Q₂ is a bond and T₂ is —OR_(c), —NR_(c)R_(d), —C(O)R_(c),—C(O)OR_(e), —S(O)₂R_(c), C₁-C₆ alkyl, or 4 to 7-memberedheterocycloalkyl, each of which is optionally substituted with one ormore -Q₃-T₃ when R_(c) or R_(d) is not H.

For example, -Q₃-T₃ is oxo.

For example, T₂ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₃-T₃ are oxo.

For example, Q₃ is a bond or unsubstituted or substituted C₁-C₃ alkyllinker.

For example, T₃ is H, halo, 4 to 7-membered heterocycloalkyl, C₁-C₃alkyl, OR_(e), COOR_(e), —S(O)₂R_(e), —NR_(e)R_(f), or —C(O)NR_(e)R_(f).

For example, one of R_(d) and R_(e) is H.

For example, Q₃ is a bond or C₁-C₃ alkyl linker and T₃ is selected fromthe group consisting of C₁-C₃ alkyl, halo, OR_(e), —S(O)₂R_(e),—NR_(e)R_(f), and —C(O)NR_(e)R_(f).

For example, R_(e) is H.

For example, R_(f) is H.

For example, R₇ is C₁-C₆ alkyl optionally substituted with one or more-Q₅-T₅.

For example, R₇ is C₃-C₈ cycloalkyl optionally substituted with one ormore -Q₅-T₅.

For example, R₇ is 4 to 7-membered heterocycloalkyl (e.g., azetidinyl,oxetanyl, thietanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl,oxazolidinyl, isoxazolidinyl, triazolidinyl, tetrahyrofuranyl,piperidinyl, 1,2,3,6-tetrahydropyridinyl, piperazinyl,tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, and morpholinyl, and thelike) optionally substituted with one or more -Q₅-T₅.

For example, R₇ is cyclopentyl.

For example, R₇ is isopropyl.

For example, R₇ is 5 to 6-membered heterocycloalkyl optionallysubstituted with one or more -Q₅-T₅.

For example, R₇ is piperidinyl optionally substituted with one -Q₅-T₅.

For example, -Q₅-T₅ is oxo.

For example, T₄ is 4 to 7-membered heterocycloalkyl or C₃-C₈ cycloalkyland one or more -Q₅-T₅ are oxo.

For example, T₅ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, C₆-C₁₀ aryl, or 4 to 7-membered heterocycloalkyl.

For example, Q₅ is a bond and T₅ is C₁-C₆ alkyl, C₃-C₈ cycloalkyl, or 4to 7-membered heterocycloalkyl.

For example, Q₅ is CO and T₅ is C₁-C₆ alkyl, C₁-C₆ alkoxyl, C₃-C₈cycloalkyl, or 4 to 7-membered heterocycloalkyl.

For example, T₅ is C₁-C₆ alkyl optionally substituted with halo,hydroxyl, cyano, C₁-C₆ alkoxyl, amino, mono-C₁-C₆ alkylamino, di-C₁-C₆alkylamino, or C₃-C₈ cycloalkyl.

For example, Q₅ is C₁-C₃ alkyl linker and T₅ is H or C₆-C₁₀ aryl.

For example, R₁₁ is H.

For example, R₅ is H, methyl, or ethyl.

For example, A⁻ is Br⁻.

For example,

For example,

is selected from the group consisting of

For example,

is selected from the group consisting of

The present invention provides the compounds of Formulae (II) and (III):

or a pharmaceutically acceptable salt or ester thereof, wherein R₁, R₂,R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, and R₁₁ are defined herein.

The present invention provides the compounds of Formulae (IIa)-(IId) and(IIIa)-(IIId):

or a pharmaceutically acceptable salt or ester thereof, wherein R₁, R₂,R₃, R₄, R₅, R₆, R₇, R₈, and R₁₁ are defined herein.

Representative compounds of the present invention include compoundslisted in Tables 1-6 below. In the tables below, each occurrence of

should be construed as

TABLE 1 Compound Number Structure A-1

A-2

A-3

A-4

A-5

A-6

A-7

A-8

A-9

A-10

A-11

A-12

A-13

A-14

A-15

A-16

A-17

A-18

A-19

A-20

A-21

A-22

A-23

A-24

A-25

A-26

A-27

A-28

A-29

A-30

A-31

A-32

A-33

A-34

A-35

A-36

A-37

A-38

A-39

A-40

A-41

A-42

A-43

A-44

A-45

A-46

A-47

A-48

A-49

A-50

A-51

A-52

A-53

A-54

A-55

A-56

A-57

A-58

A-59

A-60

A-61

A-62

A-63

A-64

A-65

A-66

A-67

A-68

A-69

A-70

A-71

A-72

A-73

A-74

A-75

A-76

A-77

A-78

A-91

A-92

A-93

A-94

A-95

A-96

A-97

A-98

A-99

A-100

A-101

A-102

A-103

A-104

A-105

A-106

A-107

A-108

A-109

A-110

A-125

A-126

TABLE 2 Compound Number Structure B-1

B-2

B-3

B-4

B-5

B-6

B-7

B-8

B-9

B-10

B-11

B-12

B-13

B-14

B-15

B-16

B-17

B-18

B-19

B-20

B-21

B-22

B-23

B-24

B-25

B-26

B-27

B-28

B-29

B-30

B-31

B-32

B-33

B-34

B-35

B-36

B-37

B-38

B-39

B-40

B-41

B-42

B-43

B-44

B-45

B-46

B-47

B-48

B-49

B-50

B-51

B-52

B-53

B-54

B-55

B-56

B-57

B-58

B-59

B-60

B-61

B-62

B-63

B-64

B-65

B-66

B-67

B-68

B-69

B-70

B-71

B-72

B-73

B-74

B-75

B-76

B-77

B-78

B-79

B-80

B-81

B-82

B-83

B-84

B-85

B-86

B-87

B-88

B-89

B-90

B-91

B-92

B-93

B-94

B-95

B-96

B-97

B-98

B-99

B-100

B-101

B-102

B-103

B-104

B-105

B-106

B-107

B-108

B-109

B-110

B-111

B-112

B-113

B-114

B-115

B-116

B-117

B-118

B-119

B-120

B-121

B-122

B-123

B-124

B-125

B-126

B-127

B-128

B-129

B-130

B-131

B-132

B-133

B-134

B-135

B-136

B-137

B-138

B-139

B-140

B-141

B-142

B-143

B-144

B-145

B-146

B-147

B-148

B-151

B-152

B-153

B-154

B-155

B-156

B-164

TABLE 3 Compound Number Structure C-1.

C-2.

C-3.

C-4.

C-5.

C-6.

C-7.

C-8.

C-9.

C-10.

C-11.

C-12.

C-13.

C-14.

C-15.

C-16.

C-17.

C-18.

C-19.

C-20.

C-21.

C-22.

C-23.

C-24.

C-25.

C-26.

C-27.

C-28.

C-29.

C-30.

C-31.

C-32.

C-33.

C-34.

C-35.

TABLE 4 Com- pound Number Structure D-1.

D-2.

D-3.

D-4.

D-5.

D-6.

D-7.

D-8.

D-9.

D-10.

D-11.

D-12.

D-13.

D-14.

D-15.

D-16.

D-17.

D-18.

D-19.

D-20.

D-21.

D-22.

D-23.

D-24.

D-25.

D-26.

D-27.

D-28.

D-29.

D-30.

D-31.

D-32.

D-33.

D-34.

D-35.

D-36.

D-37.

D-38.

D-39.

D-40.

D-41.

D-42.

D-43.

D-44.

D-45.

D-46.

D-47.

D-48.

D-49.

D-50.

D-51.

D-52.

D-53.

D-54.

D-55.

D-56.

D-57.

D-58.

D-59.

D-60.

D-61.

D-62.

D-63.

D-64.

D-65.

D-66.

D-67.

D-68.

D-69.

D-70.

D-71.

D-72.

D-73.

D-74.

D-75.

D-76.

D-77.

D-78.

D-79.

D-80.

D-81.

D-82.

D-83.

D-84.

D-85.

D-86.

D-87.

D-88.

D-89.

D-90.

D-91.

D-93

D-94

D-95

D-96

D-97

D-98

TABLE 5 Compound Number Structure E-1.

E-2.

TABLE 6 Compound Number Structure F-1.

F-2.

As used herein, “alkyl”, “C₁, C₂, C₃, C₄, C₅ or C₆ alkyl” or “C₁-C₆alkyl” is intended to include C₁, C₂, C₃, C₄, C₅ or C₆ straight chain(linear) saturated aliphatic hydrocarbon groups and C₃, C₄, C₅ or C₆branched saturated aliphatic hydrocarbon groups. For example, C₁-C₆alkyl is intended to include C₁, C₂, C₃, C₄, C₅ and C₆ alkyl groups.Examples of alkyl include, moieties having from one to six carbon atoms,such as, but not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl,s-butyl, t-butyl, n-pentyl, s-pentyl or n-hexyl.

In certain embodiments, a straight chain or branched alkyl has six orfewer carbon atoms (e.g., C₁-C₆ for straight chain, C₃-C₆ for branchedchain), and in another embodiment, a straight chain or branched alkylhas four or fewer carbon atoms.

As used herein, the term “cycloalkyl” refers to a saturated orunsaturated nonaromatic hydrocarbon mono- or multi-ring system having 3to 30 carbon atoms (e.g., C₃-C₁₀). Examples of cycloalkyl include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, andadamantyl. The term “heterocycloalkyl” refers to a saturated orunsaturated nonaromatic 3-8 membered monocyclic, 8-12 membered bicyclic,or 11-14 membered tricyclic ring system having one or more heteroatoms(such as O, N, S, or Se), unless specified otherwise. Examples ofheterocycloalkyl groups include, but are not limited to, piperidinyl,piperazinyl, pyrrolidinyl, dioxanyl, tetrahydrofuranyl, isoindolinyl,indolinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl, isoxazolidinyl,triazolidinyl, tetrahyrofuranyl, oxiranyl, azetidinyl, oxetanyl,thietanyl, 1,2,3,6-tetrahydropyridinyl, tetrahydropyranyl,dihydropyranyl, pyranyl, morpholinyl, and the like.

The term “optionally substituted alkyl” refers to unsubstituted alkyl oralkyl having designated substituents replacing one or more hydrogenatoms on one or more carbons of the hydrocarbon backbone. Suchsubstituents can include, for example, alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, amino(including alkylamino, dialkylamino, arylamino, diarylamino andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

An “arylalkyl” or an “aralkyl” moiety is an alkyl substituted with anaryl (e.g., phenylmethyl (benzyl)). An “alkylaryl” moiety is an arylsubstituted with an alkyl (e.g., methylphenyl).

As used herein, “alkyl linker” is intended to include C₁, C₂, C₃, C₄, C₅or C₆ straight chain (linear) saturated divalent aliphatic hydrocarbongroups and C₃, C₄, C₅ or C₆ branched saturated aliphatic hydrocarbongroups. For example, C₁-C₆ alkyl linker is intended to include C₁, C₂,C₃, C₄, C₅ and C₆ alkyl linker groups. Examples of alkyl linker include,moieties having from one to six carbon atoms, such as, but not limitedto, methyl (—CH₂—), ethyl (—CH₂CH₂—), n-propyl (—CH₂CH₂CH₂—), i-propyl(—CHCH₃CH₂—), n-butyl (—CH₂CH₂CH₂CH₂—), s-butyl (—CHCH₃CH₂CH₂—), i-butyl(—C(CH₃)₂CH₂—), n-pentyl (—CH₂CH₂CH₂CH₂CH₂—), s-pentyl(—CHCH₃CH₂CH₂CH₂—) or n-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₂—).

“Alkenyl” includes unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but that contain atleast one double bond. For example, the term “alkenyl” includes straightchain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl,hexenyl, heptenyl, octenyl, nonenyl, decenyl), and branched alkenylgroups. In certain embodiments, a straight chain or branched alkenylgroup has six or fewer carbon atoms in its backbone (e.g., C₂-C₆ forstraight chain, C₃-C₆ for branched chain). The term “C₂-C₆” includesalkenyl groups containing two to six carbon atoms. The term “C₃-C₆”includes alkenyl groups containing three to six carbon atoms.

The term “optionally substituted alkenyl” refers to unsubstitutedalkenyl or alkenyl having designated substituents replacing one or morehydrogen atoms on one or more hydrocarbon backbone carbon atoms. Suchsubstituents can include, for example, alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, amino(including alkylamino, dialkylamino, arylamino, diarylamino andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

“Alkynyl” includes unsaturated aliphatic groups analogous in length andpossible substitution to the alkyls described above, but which containat least one triple bond. For example, “alkynyl” includes straight chainalkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl,heptynyl, octynyl, nonynyl, decynyl), and branched alkynyl groups. Incertain embodiments, a straight chain or branched alkynyl group has sixor fewer carbon atoms in its backbone (e.g., C₂-C₆ for straight chain,C₃-C₆ for branched chain). The term “C₂-C₆” includes alkynyl groupscontaining two to six carbon atoms. The term “C₃-C₆” includes alkynylgroups containing three to six carbon atoms.

The term “optionally substituted alkynyl” refers to unsubstitutedalkynyl or alkynyl having designated substituents replacing one or morehydrogen atoms on one or more hydrocarbon backbone carbon atoms. Suchsubstituents can include, for example, alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, amino(including alkylamino, dialkylamino, acylamino, diarylamino andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl,alkylthio, arylthio, thiocarboxylate, sulfates, alkylsulfinyl,sulfonato, sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.

Other optionally substituted moieties (such as optionally substitutedcycloalkyl, heterocycloalkyl, aryl, or heteroaryl) include both theunsubstituted moieties and the moieties having one or more of thedesignated substituents. For example, substituted heterocycloalkylincludes those substituted with one or more alkyl groups, such as2,2,6,6-tetramethyl-piperidinyl and2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridinyl.

“Aryl” includes groups with aromaticity, including “conjugated,” ormulticyclic systems with at least one aromatic ring and do not containany heteroatom in the ring structure. Examples include phenyl, benzyl,1,2,3,4-tetrahydronaphthalenyl, etc.

“Heteroaryl” groups are aryl groups, as defined above, except havingfrom one to four heteroatoms in the ring structure, and may also bereferred to as “aryl heterocycles” or “heteroaromatics.” As used herein,the term “heteroaryl” is intended to include a stable 5-, 6-, or7-membered monocyclic or 7-, 8-, 9-, 10-, 11- or 12-membered bicyclicaromatic heterocyclic ring which consists of carbon atoms and one ormore heteroatoms, e.g., 1 or 1-2 or 1-3 or 1-4 or 1-5 or 1-6heteroatoms, or e.g. 1, 2, 3, 4, 5, or 6 heteroatoms, independentlyselected from the group consisting of nitrogen, oxygen and sulfur. Thenitrogen atom may be substituted or unsubstituted (i.e., N or NR whereinR is H or other substituents, as defined). The nitrogen and sulfurheteroatoms may optionally be oxidized (i.e., N→O and S(O)_(p), wherep=1 or 2). It is to be noted that total number of S and O atoms in thearomatic heterocycle is not more than 1.

Examples of heteroaryl groups include pyrrole, furan, thiophene,thiazole, isothiazole, imidazole, triazole, tetrazole, pyrazole,oxazole, isoxazole, pyridine, pyrazine, pyridazine, pyrimidine, and thelike.

Furthermore, the terms “aryl” and “heteroaryl” include multicyclic aryland heteroaryl groups, e.g., tricyclic, bicyclic, e.g., naphthalene,benzoxazole, benzodioxazole, benzothiazole, benzoimidazole,benzothiophene, methylenedioxyphenyl, quinoline, isoquinoline,naphthrydine, indole, benzofuran, purine, benzofuran, deazapurine,indolizine.

In the case of multicyclic aromatic rings, only one of the rings needsto be aromatic (e.g., 2,3-dihydroindole), although all of the rings maybe aromatic (e.g., quinoline). The second ring can also be fused orbridged.

The cycloalkyl, heterocycloalkyl, aryl, or heteroaryl ring can besubstituted at one or more ring positions (e.g., the ring-forming carbonor heteroatom such as N) with such substituents as described above, forexample, alkyl, alkenyl, akynyl, halogen, hydroxyl, alkoxy,alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkylaminocarbonyl,aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylthiocarbonyl, phosphate, phosphonato, phosphinato, amino (includingalkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moiety. Aryl and heteroarylgroups can also be fused or bridged with alicyclic or heterocyclicrings, which are not aromatic so as to form a multicyclic system (e.g.,tetralin, methylenedioxyphenyl).

As used herein, “carbocycle” or “carbocyclic ring” is intended toinclude any stable monocyclic, bicyclic or tricyclic ring having thespecified number of carbons, any of which may be saturated, unsaturated,or aromatic. Carbocycle includes cycloalkyl and aryl. For example, aC₃-C₁₄ carbocycle is intended to include a monocyclic, bicyclic ortricyclic ring having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbonatoms. Examples of carbocycles include, but are not limited to,cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl,cyclohexyl, cycloheptenyl, cycloheptyl, cycloheptenyl, adamantyl,cyclooctyl, cyclooctenyl, cyclooctadienyl, fluorenyl, phenyl, naphthyl,indanyl, adamantyl and tetrahydronaphthyl. Bridged rings are alsoincluded in the definition of carbocycle, including, for example,[3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane and[2.2.2]bicyclooctane. A bridged ring occurs when one or more carbonatoms link two non-adjacent carbon atoms. In one embodiment, bridgerings are one or two carbon atoms. It is noted that a bridge alwaysconverts a monocyclic ring into a tricyclic ring. When a ring isbridged, the substituents recited for the ring may also be present onthe bridge. Fused (e.g., naphthyl, tetrahydronaphthyl) and spiro ringsare also included.

As used herein, “heterocycle” or “heterocyclic group” includes any ringstructure (saturated, unsaturated, or aromatic) which contains at leastone ring heteroatom (e.g., N, O or S). Heterocycle includesheterocycloalkyl and heteroaryl. Examples of heterocycles include, butare not limited to, morpholine, pyrrolidine, tetrahydrothiophene,piperidine, piperazine oxetane, pyran, tetrahydropyran, azetidine, andtetrahydrofuran.

Examples of heterocyclic groups include, but are not limited to,acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl,benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl,benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl,chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl,imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl,indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl,isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl,isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl,naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl,1,2,4-oxadiazol5(4H)-one, oxazolidinyl, oxazolyl, oxindolyl,pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl,piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl,pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl,pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl,pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl,quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl,1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthenyl.

The term “substituted,” as used herein, means that any one or morehydrogen atoms on the designated atom is replaced with a selection fromthe indicated groups, provided that the designated atom's normal valencyis not exceeded, and that the substitution results in a stable compound.When a substituent is oxo or keto (i.e., ═O), then 2 hydrogen atoms onthe atom are replaced. Keto substituents are not present on aromaticmoieties. Ring double bonds, as used herein, are double bonds that areformed between two adjacent ring atoms (e.g., C═C, C═N or N═N). “Stablecompound” and “stable structure” are meant to indicate a compound thatis sufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

When a bond to a substituent is shown to cross a bond connecting twoatoms in a ring, then such substituent may be bonded to any atom in thering. When a substituent is listed without indicating the atom via whichsuch substituent is bonded to the rest of the compound of a givenformula, then such substituent may be bonded via any atom in suchformula. Combinations of substituents and/or variables are permissible,but only if such combinations result in stable compounds.

When any variable (e.g., R₁) occurs more than one time in anyconstituent or formula for a compound, its definition at each occurrenceis independent of its definition at every other occurrence. Thus, forexample, if a group is shown to be substituted with 0-2 R₁ moieties,then the group may optionally be substituted with up to two R₁ moietiesand R₁ at each occurrence is selected independently from the definitionof R₁. Also, combinations of substituents and/or variables arepermissible, but only if such combinations result in stable compounds.

The term “hydroxy” or “hydroxyl” includes groups with an —OH or —O⁻.

As used herein, “halo” or “halogen” refers to fluoro, chloro, bromo andiodo. The term “perhalogenated” generally refers to a moiety wherein allhydrogen atoms are replaced by halogen atoms. The term “haloalkyl” or“haloalkoxyl” refers to an alkyl or alkoxyl substituted with one or morehalogen atoms.

The term “carbonyl” includes compounds and moieties which contain acarbon connected with a double bond to an oxygen atom. Examples ofmoieties containing a carbonyl include, but are not limited to,aldehydes, ketones, carboxylic acids, amides, esters, anhydrides, etc.

The term “carboxyl” refers to —COOH or its C₁-C₆ alkyl ester.

“Acyl” includes moieties that contain the acyl radical (R—C(O)—) or acarbonyl group. “Substituted acyl” includes acyl groups where one ormore of the hydrogen atoms are replaced by, for example, alkyl groups,alkynyl groups, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, amino (including alkylamino, dialkylamino,arylamino, diarylamino and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

“Aroyl” includes moieties with an aryl or heteroaromatic moiety bound toa carbonyl group. Examples of aroyl groups include phenylcarboxy,naphthyl carboxy, etc.

“Alkoxyalkyl,” “alkylaminoalkyl,” and “thioalkoxyalkyl” include alkylgroups, as described above, wherein oxygen, nitrogen, or sulfur atomsreplace one or more hydrocarbon backbone carbon atoms.

The term “alkoxy” or “alkoxyl” includes substituted and unsubstitutedalkyl, alkenyl and alkynyl groups covalently linked to an oxygen atom.Examples of alkoxy groups or alkoxyl radicals include, but are notlimited to, methoxy, ethoxy, isopropyloxy, propoxy, butoxy and pentoxygroups. Examples of substituted alkoxy groups include halogenated alkoxygroups. The alkoxy groups can be substituted with groups such asalkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, amino (including alkylamino, dialkylamino,arylamino, diarylamino, and alkylarylamino), acylamino (includingalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino,imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates,alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moieties. Examples of halogen substituted alkoxygroups include, but are not limited to, fluoromethoxy, difluoromethoxy,trifluoromethoxy, chloromethoxy, dichloromethoxy and trichloromethoxy.

The term “ether” or “alkoxy” includes compounds or moieties whichcontain an oxygen bonded to two carbon atoms or heteroatoms. Forexample, the term includes “alkoxyalkyl,” which refers to an alkyl,alkenyl, or alkynyl group covalently bonded to an oxygen atom which iscovalently bonded to an alkyl group.

The term “ester” includes compounds or moieties which contain a carbonor a heteroatom bound to an oxygen atom which is bonded to the carbon ofa carbonyl group. The term “ester” includes alkoxycarboxy groups such asmethoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl,pentoxycarbonyl, etc.

The term “thioalkyl” includes compounds or moieties which contain analkyl group connected with a sulfur atom. The thioalkyl groups can besubstituted with groups such as alkyl, alkenyl, alkynyl, halogen,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,aryloxycarbonyloxy, carboxylate, carboxyacid, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, amino (includingalkylamino, dialkylamino, arylamino, diarylamino and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moieties.

The term “thiocarbonyl” or “thiocarboxy” includes compounds and moietieswhich contain a carbon connected with a double bond to a sulfur atom.

The term “thioether” includes moieties which contain a sulfur atombonded to two carbon atoms or heteroatoms. Examples of thioethersinclude, but are not limited to alkthioalkyls, alkthioalkenyls, andalkthioalkynyls. The term “alkthioalkyls” include moieties with analkyl, alkenyl, or alkynyl group bonded to a sulfur atom which is bondedto an alkyl group. Similarly, the term “alkthioalkenyls” refers tomoieties wherein an alkyl, alkenyl or alkynyl group is bonded to asulfur atom which is covalently bonded to an alkenyl group; andalkthioalkynyls” refers to moieties wherein an alkyl, alkenyl or alkynylgroup is bonded to a sulfur atom which is covalently bonded to analkynyl group.

As used herein, “amine” or “amino” refers to unsubstituted orsubstituted —NH₂. “Alkylamino” includes groups of compounds whereinnitrogen of —NH₂ is bound to at least one alkyl group. Examples ofalkylamino groups include benzylamino, methylamino, ethylamino,phenethylamino, etc. “Dialkylamino” includes groups wherein the nitrogenof —NH₂ is bound to at least two additional alkyl groups. Examples ofdialkylamino groups include, but are not limited to, dimethylamino anddiethylamine. “Arylamino” and “diarylamino” include groups wherein thenitrogen is bound to at least one or two aryl groups, respectively.“Aminorcyl” and “aminoaryloxy” refer to aryl and aryloxy substitutedwith amino. “Alkylarylamino,” “alkylaminoaryl” or “arylaminoalkyl”refers to an amino group which is bound to at least one alkyl group andat least one aryl group. “Alkaminoalkyl” refers to an alkyl, alkenyl, oralkynyl group bound to a nitrogen atom which is also bound to an alkylgroup. “Acylamino” includes groups wherein nitrogen is bound to an acylgroup. Examples of acylamino include, but are not limited to,alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido groups.

The term “amide” or “aminocarboxy” includes compounds or moieties thatcontain a nitrogen atom that is bound to the carbon of a carbonyl or athiocarbonyl group. The term includes “alkaminocarboxy” groups thatinclude alkyl, alkenyl or alkynyl groups bound to an amino group whichis bound to the carbon of a carbonyl or thiocarbonyl group. It alsoincludes “arylaminocarboxy” groups that include aryl or heteroarylmoieties bound to an amino group that is bound to the carbon of acarbonyl or thiocarbonyl group. The terms “alkylaminocarboxy”,“alkenylaminocarboxy”, “alkynylaminocarboxy” and “arylaminocarboxy”include moieties wherein alkyl, alkenyl, alkynyl and aryl moieties,respectively, are bound to a nitrogen atom which is in turn bound to thecarbon of a carbonyl group. Amides can be substituted with substituentssuch as straight chain alkyl, branched alkyl, cycloalkyl, aryl,heteroaryl or heterocycle. Substituents on amide groups may be furthersubstituted.

Compounds of the present invention that contain nitrogens can beconverted to N-oxides by treatment with an oxidizing agent (e.g.,3-chloroperoxybenzoic acid (mCPBA) and/or hydrogen peroxides) to affordother compounds of the present invention. Thus, all shown and claimednitrogen-containing compounds are considered, when allowed by valencyand structure, to include both the compound as shown and its N-oxidederivative (which can be designated as N→O or N⁺—O⁻). Furthermore, inother instances, the nitrogens in the compounds of the present inventioncan be converted to N-hydroxy or N-alkoxy compounds. For example,N-hydroxy compounds can be prepared by oxidation of the parent amine byan oxidizing agent such as m-CPBA. All shown and claimednitrogen-containing compounds are also considered, when allowed byvalency and structure, to cover both the compound as shown and itsN-hydroxy (i.e., N—OH) and N-alkoxy (i.e., N—OR, wherein R issubstituted or unsubstituted C₁-C₆ alkyl, C₁-C₆ alkenyl, C₁-C₆ alkynyl,3-14-membered carbocycle or 3-14-membered heterocycle) derivatives.

In the present specification, the structural formula of the compoundrepresents a certain isomer for convenience in some cases, but thepresent invention includes all isomers, such as geometrical isomers,optical isomers based on an asymmetrical carbon, stereoisomers,tautomers, and the like. In addition, a crystal polymorphism may bepresent for the compounds represented by the formula. It is noted thatany crystal form, crystal form mixture, or anhydride or hydrate thereofis included in the scope of the present invention. Furthermore,so-called metabolite which is produced by degradation of the presentcompound in vivo is included in the scope of the present invention.

“Isomerism” means compounds that have identical molecular formulae butdiffer in the sequence of bonding of their atoms or in the arrangementof their atoms in space. Isomers that differ in the arrangement of theiratoms in space are termed “stereoisomers.” Stereoisomers that are notmirror images of one another are termed “diastereoisomers,” andstereoisomers that are non-superimposable mirror images of each otherare termed “enantiomers” or sometimes optical isomers. A mixturecontaining equal amounts of individual enantiomeric forms of oppositechirality is termed a “racemic mixture.”

A carbon atom bonded to four nonidentical substituents is termed a“chiral center.”

“Chiral isomer” means a compound with at least one chiral center.Compounds with more than one chiral center may exist either as anindividual diastereomer or as a mixture of diastereomers, termed“diastereomeric mixture.” When one chiral center is present, astereoisomer may be characterized by the absolute configuration (R or S)of that chiral center. Absolute configuration refers to the arrangementin space of the substituents attached to the chiral center. Thesubstituents attached to the chiral center under consideration areranked in accordance with the Sequence Rule of Cahn, Ingold and Prelog.(Cahn et al., Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahnet al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J. Chem. Soc. 1951(London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn, J. Chem.Educ. 1964, 41, 116).

“Geometric isomer” means the diastereomers that owe their existence tohindered rotation about double bonds or a cycloalkyl linker (e.g.,1,3-cylcobutyl). These configurations are differentiated in their namesby the prefixes cis and trans, or Z and E, which indicate that thegroups are on the same or opposite side of the double bond in themolecule according to the Cahn-Ingold-Prelog rules.

It is to be understood that the compounds of the present invention maybe depicted as different chiral isomers or geometric isomers. It shouldalso be understood that when compounds have chiral isomeric or geometricisomeric forms, all isomeric forms are intended to be included in thescope of the present invention, and the naming of the compounds does notexclude any isomeric forms.

Furthermore, the structures and other compounds discussed in thisinvention include all atropic isomers thereof “Atropic isomers” are atype of stereoisomer in which the atoms of two isomers are arrangeddifferently in space. Atropic isomers owe their existence to arestricted rotation caused by hindrance of rotation of large groupsabout a central bond. Such atropic isomers typically exist as a mixture,however as a result of recent advances in chromatography techniques, ithas been possible to separate mixtures of two atropic isomers in selectcases.

“Tautomer” is one of two or more structural isomers that exist inequilibrium and is readily converted from one isomeric form to another.This conversion results in the formal migration of a hydrogen atomaccompanied by a switch of adjacent conjugated double bonds. Tautomersexist as a mixture of a tautomeric set in solution. In solutions wheretautomerization is possible, a chemical equilibrium of the tautomerswill be reached. The exact ratio of the tautomers depends on severalfactors, including temperature, solvent and pH. The concept of tautomersthat are interconvertable by tautomerizations is called tautomerism.

Of the various types of tautomerism that are possible, two are commonlyobserved. In keto-enol tautomerism a simultaneous shift of electrons anda hydrogen atom occurs. Ring-chain tautomerism arises as a result of thealdehyde group (—CHO) in a sugar chain molecule reacting with one of thehydroxy groups (—OH) in the same molecule to give it a cyclic(ring-shaped) form as exhibited by glucose.

Common tautomeric pairs are: ketone-enol, amide-nitrile, lactam-lactim,amide-imidic acid tautomerism in heterocyclic rings (e.g., innucleobases such as guanine, thymine and cytosine), amine-enamine andenamine-enamine. An example of keto-enol equilibria is betweenpyridin-2(1H)-one's and the corresponding pyridin-2-ol's, as shownbelow.

It is to be understood that the compounds of the present invention maybe depicted as different tautomers. It should also be understood thatwhen compounds have tautomeric forms, all tautomeric forms are intendedto be included in the scope of the present invention, and the naming ofthe compounds does not exclude any tautomer form.

The term “crystal polymorphs”, “polymorphs” or “crystal forms” meanscrystal structures in which a compound (or a salt or solvate thereof)can crystallize in different crystal packing arrangements, all of whichhave the same elemental composition. Different crystal forms usuallyhave different X-ray diffraction patterns, infrared spectral, meltingpoints, density hardness, crystal shape, optical and electricalproperties, stability and solubility. Recrystallization solvent, rate ofcrystallization, storage temperature, and other factors may cause onecrystal form to dominate. Crystal polymorphs of the compounds can beprepared by crystallization under different conditions.

The compounds of Formula (I), (Ia), or (Ib) include the compoundsthemselves, as well as their salts, their solvates, and their prodrugs,if applicable. A salt, for example, can be formed between an anion and apositively charged group (e.g., amino) on a substituted 6,5-fusedbicyclic heteroaryl compound. Suitable anions include chloride, bromide,iodide, sulfate, bisulfate, sulfamate, nitrate, phosphate, citrate,methanesulfonate, trifluoroacetate, glutamate, glucuronate, glutarate,malate, maleate, succinate, fumarate, tartrate, tosylate, salicylate,lactate, naphthalenesulfonate, and acetate (e.g., trifluroacetate). Theterm “pharmaceutically acceptable anion” refers to an anion suitable forforming a pharmaceutically acceptable salt. Likewise, a salt can also beformed between a cation and a negatively charged group (e.g.,carboxylate) on a substituted 6,5-fused bicyclic heteroaryl compound.Suitable cations include sodium ion, potassium ion, magnesium ion,calcium ion, and an ammonium cation such as tetramethylammonium ion. Thesubstituted 6,5-fused bicyclic heteroaryl compounds also include thosesalts containing quaternary nitrogen atoms. Examples of prodrugs includeesters and other pharmaceutically acceptable derivatives, which, uponadministration to a subject, are capable of providing active substituted6,5-fused bicyclic heteroaryl compounds.

Additionally, the compounds of the present invention, for example, thesalts of the compounds, can exist in either hydrated or unhydrated (theanhydrous) form or as solvates with other solvent molecules. Nonlimitingexamples of hydrates include monohydrates, dihydrates, etc. Nonlimitingexamples of solvates include ethanol solvates, acetone solvates, etc.

“Solvate” means solvent addition forms that contain eitherstoichiometric or non stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate; and if the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one molecule of the substance inwhich the water retains its molecular state as H₂O.

As used herein, the term “analog” refers to a chemical compound that isstructurally similar to another but differs slightly in composition (asin the replacement of one atom by an atom of a different element or inthe presence of a particular functional group, or the replacement of onefunctional group by another functional group). Thus, an analog is acompound that is similar or comparable in function and appearance, butnot in structure or origin to the reference compound.

As defined herein, the term “derivative” refers to compounds that have acommon core structure, and are substituted with various groups asdescribed herein. For example, all of the compounds represented byFormula (I) are substituted 6,5-fused bicyclic heteroaryl compounds, andhave Formula (I) as a common core.

The term “bioisostere” refers to a compound resulting from the exchangeof an atom or of a group of atoms with another, broadly similar, atom orgroup of atoms. The objective of a bioisosteric replacement is to createa new compound with similar biological properties to the parentcompound. The bioisosteric replacement may be physicochemically ortopologically based. Examples of carboxylic acid bioisosteres include,but are not limited to, acyl sulfonimides, tetrazoles, sulfonates andphosphonates. See, e.g., Patani and LaVoie, Chem. Rev. 96, 3147-3176,1996.

The present invention is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include tritium anddeuterium, and isotopes of carbon include C-13 and C-14.

2. Synthesis of Substituted 6,5-fused Bicyclic Heteroaryl Compounds

The present invention provides methods for the synthesis of thecompounds of Formulae (I), (Ia), and (Ib). The present invention alsoprovides detailed methods for the synthesis of various disclosedcompounds of the present invention according to the following schemes asshown in the Examples.

Throughout the description, where compositions are described as having,including, or comprising specific components, it is contemplated thatcompositions also consist essentially of, or consist of, the recitedcomponents. Similarly, where methods or processes are described ashaving, including, or comprising specific process steps, the processesalso consist essentially of, or consist of, the recited processingsteps. Further, it should be understood that the order of steps or orderfor performing certain actions is immaterial so long as the inventionremains operable. Moreover, two or more steps or actions can beconducted simultaneously.

The synthetic processes of the invention can tolerate a wide variety offunctional groups, therefore various substituted starting materials canbe used. The processes generally provide the desired final compound ator near the end of the overall process, although it may be desirable incertain instances to further convert the compound to a pharmaceuticallyacceptable salt, ester, or prodrug thereof.

Compounds of the present invention can be prepared in a variety of waysusing commercially available starting materials, compounds known in theliterature, or from readily prepared intermediates, by employingstandard synthetic methods and procedures either known to those skilledin the art, or which will be apparent to the skilled artisan in light ofthe teachings herein. Standard synthetic methods and procedures for thepreparation of organic molecules and functional group transformationsand manipulations can be obtained from the relevant scientificliterature or from standard textbooks in the field. Although not limitedto any one or several sources, classic texts such as Smith, M. B.,March, J., March's Advanced Organic Chemistry: Reactions, Mechanisms,and Structure, 5th edition, John Wiley & Sons: New York, 2001; Greene,T. W., Wuts, P. G. M., Protective Groups in Organic Synthesis, 3^(rd)edition, John Wiley & Sons: New York, 1999; R. Larock, ComprehensiveOrganic Transformations, VCH Publishers (1989); L. Fieser and M. Fieser,Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons(1994); and L. Paquette, ed., Encyclopedia of Reagents for OrganicSynthesis, John Wiley and Sons (1995), incorporated by reference herein,are useful and recognized reference textbooks of organic synthesis knownto those in the art. The following descriptions of synthetic methods aredesigned to illustrate, but not to limit, general procedures for thepreparation of compounds of the present invention.

Compounds of the present invention can be conveniently prepared by avariety of methods familiar to those skilled in the art. The compoundsof this invention with Formulae (I), (Ia), and (Ib) may be preparedaccording to the procedures illustrated in Schemes 1-4 below, fromcommercially available starting materials or starting materials whichcan be prepared using literature procedures. The R groups (such as R₆,R₇, and R₁₁) in Schemes 1-4 are as defined in Formula (I), (Ia), or(Ib), unless otherwise specified.

One of ordinary skill in the art will note that, during the reactionsequences and synthetic schemes described herein, the order of certainsteps may be changed, such as the introduction and removal of protectinggroups.

One of ordinary skill in the art will recognize that certain groups mayrequire protection from the reaction conditions via the use ofprotecting groups. Protecting groups may also be used to differentiatesimilar functional groups in molecules. A list of protecting groups andhow to introduce and remove these groups can be found in Greene, T. W.,Wuts, P. G. M., Protective Groups in Organic Synthesis, 3^(rd) edition,John Wiley & Sons: New York, 1999.

Preferred protecting groups include, but are not limited to:

For the hydroxyl moiety: TBS, benzyl, THP, Ac

For carboxylic acids: benzyl ester, methyl ester, ethyl ester, allylester

For amines: Cbz, BOC, DMB

For diols: Ac (×2) TBS (×2), or when taken together acetonides

For thiols: Ac

For benzimidazoles: SEM, benzyl, PMB, DMB

For aldehydes: di-alkyl acetals such as dimethoxy acetal or diethylacetyl.

In the reaction schemes described herein, multiple stereoisomers may beproduced. When no particular stereoisomer is indicated, it is understoodto mean all possible stereoisomers that could be produced from thereaction. A person of ordinary skill in the art will recognize that thereactions can be optimized to give one isomer preferentially, or newschemes may be devised to produce a single isomer. If mixtures areproduced, techniques such as preparative thin layer chromatography,preparative HPLC, preparative chiral HPLC, or preparative SFC may beused to separate the isomers.

The following abbreviations are used throughout the specification andare defined below:

AA ammonium acetate

Ac acetyl

ACN acetonitrile

AcOH acetic acid

atm atmosphere

Bn benzyl

BOC tert-butoxy carbonyl

BOP (benzotriazol-1-yloxy)tris(dimethylamino)phosphoniumhexafluorophosphate

Cbz benzyloxy carbonyl

COMU(1-Cyano-2-ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbeniumhexafluorophosphate

d days

DBU 1,8-diazabicyclo[5.4.0]undec-7-ene

DCE 1,2 dichloroethane

DCM dichloromethane

DEAD Diethyl azodicarboxylate

DIAD Diisopropyl azodicarboxylate

DiBAL-H diisobutyl alumininium hydride

DIPEA N,N-diisopropylethylamine (Hunig's base)

DMAP N,N-dimethyl-4-aminopyridine

DMB 2,4 dimethoxy benzyl

DMF dimethylformamide

DMSO Dimethyl sulfoxide

DPPA Diphenylphosphonic azide

EA or EtOAc Ethyl acetate

EDC or EDCI N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide

ELS Evaporative Light Scattering

ESI− Electrospray negative mode

ESI+ Electrospray positive mode

Et₂O diethyl ether

Et₃N or TEA triethylamine

EtOH ethanol

FA formic acid

FC Flash chromatogrpahy

h hours

H₂O water

HATU O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate

HCl hydrochloric acid

HOAT 1-Hydroxy-7-azabenzotriazole

HOBt 1-Hydroxybenzotriazole

HO-Su N-Hydroxysuccinimide

HPLC High performance liquid chromatography

KHMDs Potassium hexamethyldisilazide

LC/MS or LC-MS liquid chromatography mass spectrum

LDA Lithium diisiopropylamide

LG leaving group

LiHMDs Lithium hexamethyldisilazide

M Molar

m/z mass/charge ratio

m-CPBA meta-chloroperbenzoic acid

MeCN Acetonitrile

MeOD d₄-methanol

MeOH methanol

MgSO₄ magnesium sulfate

min minutes

MS Mass Spectrometry

Ms Mesyl

MS mass spectrum

MsCl Mesyl chloride

MsO Mesylate

MWI microwave irradiation

Na₂CO₃ sodium carbonate

NaHCO₃ sodium bicarbonate

NaHMDs Sodium hexamethyldisilazide

NaOH sodium hydroxide

NIS N-iodosuccinimide

NMR Nuclear Magnetic Resonance

o/n or O/N overnight

PE Petroleum Ether

PG protecting group

PMB para methoxybenzyl

PPAA 1-Propanephosphonic acid cyclic anhydride

ppm parts per million

prep HPLC preparative High performance liquid chromatography

prep TLC preparative thin layer chromatography

p-TsOH para-toluenesulfonic acid

PYBOP (Benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate

rt or RT room temperature

SEM 2-(Trimethylsilyl)ethoxymethyl

SEMCl (Trimethylsilyl)ethoxymethyl chloride

SFC Super critical chromatography

SGC silica gel chromatogrpahy

STAB Sodium triacetoxy borohydride

TBAF tetra-n-butylammonium fluoride

TFA trifluoroacetic acid

TfO triflate

THF tetrahydrofuran

THP tetrahydropyran

TLC thin layer chromatography

Ts tosyl

TsOH tosic acid

UV ultraviolet

Scheme 1 shows the synthesis of modified pyrazolopyridine analogsfollowing a general route that utilizes well-established chemistry.Condensation of 1H-pyrazol-3-amine with sodium(E)-1,4-diethoxy-1,4-dioxobut-2-en-2-olate in a polar solvent such aswater using a mild acid catalyst such as acetic acid can provide thehydroxyl-pyrazolopyridine (Step 1). The hydroxyl group can then beconverted to a leaving group “X” such as bromide using phosphoryltribromide at elevated temperatures in an appropriate polar solvent suchas acetonitrile to give the bromide (Step 2). Introduction of the R₇ canbe done using an appropriate R₇-LG where LG is a leaving group such asOTs or Br. Subjecting the intermediate to R₇-LG in the presence of amild base such as potassium carbonate in an appropriate polar solventsuch as acetonitrile gives the desired substituted pyrazolopyridine(Step 3). A variety of R₆ substituents can then be introduced usingstandard transition metal-based protocols that rely upon a leaving groupsuch as a bromide as a connection point or through direct SNArdisplacement of the bromide with a nucleophile. The bromide can becombined with an appropriate boronic ester derivative, in the presenceof a mild base and a palladium catalyst in a polar solvent such asdioxane/water, at elevated temperature to give the desiredpyrazolopyridine ester (Step 4). Alternatively, the bromide can becombined with a nucleophile such as an amine in the presence of a mildbase such as potassium carbonate in a polar solvent such as acetone togive the desired pyrazolopyridine ester. The ester moiety can beconverted to an amide using a standard two step protocol. The ester canbe hydrolyzed to the corresponding acid using a suitable base such assodium hydroxide in a polar solvent such as ethanol (Step 5). The acidis then subjected to a standard amide coupling reaction whereupon theappropriate amine is added along with a suitable amide coupling reagentsuch as PYBOP in a suitable solvent such as DMSO to give the desiredamide (Step 6).

Scheme 2 shows the synthesis of modified indazole analogs following ageneral route that utilizes well-established chemistry. Introduction ofa nitro group to a tolyl compound can be achieved using standardnitration conditions such as nitric acid in sulfuric acid (Step 1). Theacid can be esterified by treatment with an alkylating agent such asmethyliodide in the presence of a base such as sodium carbonate in anappropriate polar solvent such as DMF (Step 2). Reduction of the nitrogroup using an appropriate reducing agent such as iron with an acid suchas ammonium chloride in a protic solvent such as ethanol can provide ananiline (Step 3). Diazotization with an appropriate reagent such assodium nitrite in a polar solvent such as acetic acid can lead tocyclization to provide an indazole (Step 4). It will be apparent to oneskilled in the art that there are multiple ways to synthesize indazoles(J. Org. Chem. 2006, 71, 8166-8172). Introduction of the R₇ to theindazole can be done using an appropriate R₇-LG where LG is a leavinggroup such as OTs or Br. Subjecting the intermediate to R₇-LG in thepresence of a mild base such as cesium carbonate in an appropriate polarsolvent such as DMF can give the desired R₇-substituted indazole ester(Step 5). The ester moiety can be converted to an amide using a standardtwo step protocol. The ester can be hydrolyzed to the corresponding acidusing a suitable base such as sodium hydroxide in a polar solvent suchas ethanol (Step 6). The acid can then be subjecting to a standard amidecoupling reaction whereupon the appropriate amine can be added alongwith a suitable amide coupling reagent such as PYBOP in a suitablesolvent such as DMSO to give the desired amide (Step 7).

When R₆ is an appropriate group such as bromide or triflate, a varietyof substituents could then be introduced using standard transitionmetal-based protocols. For example, the bromide can be combined with anappropriate boronic ester derivative, in the presence of a mild base anda palladium catalyst in a polar solvent such as dioxane/water, atelevated temperature to give the desired indazole (Step 8).

As shown in scheme 4, a diketone can be condensed with 2-cyanoacetamidein the presence of an appropriate reagent such as piperidine acetate ina polar solvent such as ethanol to provide a cyanopyridone (Step 9).Additionally, when R₃ is H, an appropriately substituted alkynyl ketonecan be condensed with 2-cyanoacetamide in the presence of an appropriatereagent such as piperidine acetate in a polar solvent such as ethanol toprovide a cyanopyridone (Step 11). The cyano group can be reduced underappropriate conditions such as hydrogenation in the presence ofcatalytic Raney nickel in a polar solvent such as ammonium in methanolto provide the amine (Step 10).

Additionally, depending upon the nature of the R₂, R₃, R₄, and R₆ group,further chemical modification can be employed to convert each of themindependently into an alternative substituent. A representative samplingof such modifications can include hydrogenation, protecting groupremoval followed by additional amide coupling reactions, palladiumcatalyzed coupling reactions, reductive amination reactions, andalkylation reactions.

3. Methods of Treatment

The present invention provides methods for treating conditions anddiseases the course of which can be influenced by modulating themethylation status of histones or other proteins, wherein saidmethylation status is mediated at least in part by the activity of EZH2.Modulation of the methylation status of histones can in turn influencethe level of expression of target genes activated by methylation, and/ortarget genes suppressed by methylation. The method includesadministering to a subject in need of such treatment, a therapeuticallyeffective amount of a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph,solvate, or stereoisomeror thereof

The disorder in which EZH2-mediated protein methylation plays a part canbe cancer or a precancerous condition. The present invention furtherprovides the use of a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, for the preparation of a medicament useful for thetreatment of cancer.

The present invention also provides methods of protecting against adisorder in which EZH2-mediated protein methylation plays a part in asubject in need thereof by administering a therapeutically effectiveamount of compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, polymorph or solvate thereof, to asubject in need of such treatment. The disorder can be cancer. Thepresent invention also provides the use of compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,polymorph, solvate, or stereoisomeror thereof, for the preparation of amedicament useful for the prevention of a cell proliferative disorder.

The compounds of this invention can be used to modulate protein (e.g.,histone) methylation, e.g., to modulate histone methyltransferase orhistone demethylase enzyme activity. Histone methylation has beenreported to be involved in aberrant expression of certain genes incancers, and in silencing of neuronal genes in non-neuronal cells. Thecompounds described herein can be used to treat these diseases, i.e., todecreases methylation or restores methylation to roughly its level incounterpart normal cells.

In general, compounds that are methylation modulators can be used formodulating cell proliferation, generally. For example, in some casesexcessive proliferation may be reduced with agents that decreasemethylation, whereas insufficient proliferation may be stimulated withagents that increase methylation. Accordingly, diseases that may betreated by the compounds of the invention include hyperproliferativediseases, such as benign cell growth and malignant cell growth.

As used herein, a “subject in need thereof” is a subject having adisorder in which EZH2-mediated protein methylation plays a part, or asubject having an increased risk of developing such disorder relative tothe population at large. A subject in need thereof can have aprecancerous condition. Preferably, a subject in need thereof hascancer. A “subject” includes a mammal. The mammal can be e.g., anymammal, e.g., a human, primate, bird, mouse, rat, fowl, dog, cat, cow,horse, goat, camel, sheep or a pig. Preferably, the mammal is a human.

As used herein, the term “cell proliferative disorder” refers toconditions in which unregulated or abnormal growth, or both, of cellscan lead to the development of an unwanted condition or disease, whichmay or may not be cancerous. Exemplary cell proliferative disorders ofthe invention encompass a variety of conditions wherein cell division isderegulated. Exemplary cell proliferative disorder include, but are notlimited to, neoplasms, benign tumors, malignant tumors, pre-cancerousconditions, in situ tumors, encapsulated tumors, metastatic tumors,liquid tumors, solid tumors, immunological tumors, hematological tumors,cancers, carcinomas, leukemias, lymphomas, sarcomas, and rapidlydividing cells. The term “rapidly dividing cell” as used herein isdefined as any cell that divides at a rate that exceeds or is greaterthan what is expected or observed among neighboring or juxtaposed cellswithin the same tissue. A cell proliferative disorder includes aprecancer or a precancerous condition. A cell proliferative disorderincludes cancer. Preferably, the methods provided herein are used totreat or alleviate a symptom of cancer. The term “cancer” includes solidtumors, as well as, hematologic tumors and/or malignancies. A “precancercell” or “precancerous cell” is a cell manifesting a cell proliferativedisorder that is a precancer or a precancerous condition. A “cancercell” or “cancerous cell” is a cell manifesting a cell proliferativedisorder that is a cancer. Any reproducible means of measurement may beused to identify cancer cells or precancerous cells. Cancer cells orprecancerous cells can be identified by histological typing or gradingof a tissue sample (e.g., a biopsy sample). Cancer cells or precancerouscells can be identified through the use of appropriate molecularmarkers.

Exemplary non-cancerous conditions or disorders include, but are notlimited to, rheumatoid arthritis; inflammation; autoimmune disease;lymphoproliferative conditions; acromegaly; rheumatoid spondylitis;osteoarthritis; gout, other arthritic conditions; sepsis; septic shock;endotoxic shock; gram-negative sepsis; toxic shock syndrome; asthma;adult respiratory distress syndrome; chronic obstructive pulmonarydisease; chronic pulmonary inflammation; inflammatory bowel disease;Crohn's disease; psoriasis; eczema; ulcerative colitis; pancreaticfibrosis; hepatic fibrosis; acute and chronic renal disease; irritablebowel syndrome; pyresis; restenosis; cerebral malaria; stroke andischemic injury; neural trauma; Alzheimer's disease; Huntington'sdisease; Parkinson's disease; acute and chronic pain; allergic rhinitis;allergic conjunctivitis; chronic heart failure; acute coronary syndrome;cachexia; malaria; leprosy; leishmaniasis; Lyme disease; Reiter'ssyndrome; acute synovitis; muscle degeneration, bursitis; tendonitis;tenosynovitis; herniated, ruptures, or prolapsed intervertebral disksyndrome; osteopetrosis; thrombosis; restenosis; silicosis; pulmonarysarcosis; bone resorption diseases, such as osteoporosis;graft-versus-host reaction; Multiple Sclerosis; lupus; fibromyalgia;AIDS and other viral diseases such as Herpes Zoster, Herpes Simplex I orII, influenza virus and cytomegalovirus; and diabetes mellitus.

Exemplary cancers include, but are not limited to, adrenocorticalcarcinoma, AIDS-related cancers, AIDS-related lymphoma, anal cancer,anorectal cancer, cancer of the anal canal, appendix cancer, childhoodcerebellar astrocytoma, childhood cerebral astrocytoma, basal cellcarcinoma, skin cancer (non-melanoma), biliary cancer, extrahepatic bileduct cancer, intrahepatic bile duct cancer, bladder cancer, uringarybladder cancer, bone and joint cancer, osteosarcoma and malignantfibrous histiocytoma, brain cancer, brain tumor, brain stem glioma,cerebellar astrocytoma, cerebral astrocytoma/malignant glioma,ependymoma, medulloblastoma, supratentorial primitive neuroectodeimaltumors, visual pathway and hypothalamic glioma, breast cancer, bronchialadenomas/carcinoids, carcinoid tumor, gastrointestinal, nervous systemcancer, nervous system lymphoma, central nervous system cancer, centralnervous system lymphoma, cervical cancer, childhood cancers, chroniclymphocytic leukemia, chronic myelogenous leukemia, chronicmyeloproliferative disorders, colon cancer, colorectal cancer, cutaneousT-cell lymphoma, lymphoid neoplasm, mycosis fungoides, Seziary Syndrome,endometrial cancer, esophageal cancer, extracranial germ cell tumor,extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer,intraocular melanoma, retinoblastoma, gallbladder cancer, gastric(stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinalstromal tumor (GIST), germ cell tumor, ovarian germ cell tumor,gestational trophoblastic tumor glioma, head and neck cancer,hepatocellular (liver) cancer, Hodgkin lymphoma, hypopharyngeal cancer,intraocular melanoma, ocular cancer, islet cell tumors (endocrinepancreas), Kaposi Sarcoma, kidney cancer, renal cancer, kidney cancer,laryngeal cancer, acute lymphoblastic leukemia, acute myeloid leukemia,chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cellleukemia, lip and oral cavity cancer, liver cancer, lung cancer,non-small cell lung cancer, small cell lung cancer, AIDS-relatedlymphoma, non-Hodgkin lymphoma, primary central nervous system lymphoma,Waldenstram macroglobulinemia, medulloblastoma, melanoma, intraocular(eye) melanoma, merkel cell carcinoma, mesothelioma malignant,mesothelioma, metastatic squamous neck cancer, mouth cancer, cancer ofthe tongue, multiple endocrine neoplasia syndrome, mycosis fungoides,myelodysplastic syndromes, myelodysplastic/myeloproliferative diseases,chronic myelogenous leukemia, acute myeloid leukemia, multiple myeloma,chronic myeloproliferative disorders, nasopharyngeal cancer,neuroblastoma, oral cancer, oral cavity cancer, oropharyngeal cancer,ovarian cancer, ovarian epithelial cancer, ovarian low malignantpotential tumor, pancreatic cancer, islet cell pancreatic cancer,paranasal sinus and nasal cavity cancer, parathyroid cancer, penilecancer, pharyngeal cancer, pheochromocytoma, pineoblastoma andsupratentorial primitive neuroectodermal tumors, pituitary tumor, plasmacell neoplasm/multiple myeloma, pleuropulmonary blastoma, prostatecancer, rectal cancer, renal pelvis and ureter, transitional cellcancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, ewingfamily of sarcoma tumors, Kaposi Sarcoma, soft tissue sarcoma, uterinecancer, uterine sarcoma, skin cancer (non-melanoma), skin cancer(melanoma), merkel cell skin carcinoma, small intestine cancer, softtissue sarcoma, squamous cell carcinoma, stomach (gastric) cancer,supratentorial primitive neuroectodermal tumors, testicular cancer,throat cancer, thymoma, thymoma and thymic carcinoma, thyroid cancer,transitional cell cancer of the renal pelvis and ureter and otherurinary organs, gestational trophoblastic tumor, urethral cancer,endometrial uterine cancer, uterine sarcoma, uterine corpus cancer,vaginal cancer, vulvar cancer, and Wilm's Tumor.

A “cell proliferative disorder of the hematologic system” is a cellproliferative disorder involving cells of the hematologic system. A cellproliferative disorder of the hematologic system can include lymphoma,leukemia, myeloid neoplasms, mast cell neoplasms, myelodysplasia, benignmonoclonal gammopathy, lymphomatoid granulomatosis, lymphomatoidpapulosis, polycythemia vera, chronic myelocytic leukemia, agnogenicmyeloid metaplasia, and essential thrombocythemia. A cell proliferativedisorder of the hematologic system can include hyperplasia, dysplasia,and metaplasia of cells of the hematologic system. Preferably,compositions of the present invention may be used to treat a cancerselected from the group consisting of a hematologic cancer of thepresent invention or a hematologic cell proliferative disorder of thepresent invention. A hematologic cancer of the present invention caninclude multiple myeloma, lymphoma (including Hodgkin's lymphoma,non-Hodgkin's lymphoma, childhood lymphomas, and lymphomas oflymphocytic and cutaneous origin), leukemia (including childhoodleukemia, hairy-cell leukemia, acute lymphocytic leukemia, acutemyelocytic leukemia, chronic lymphocytic leukemia, chronic myelocyticleukemia, chronic myelogenous leukemia, and mast cell leukemia), myeloidneoplasms and mast cell neoplasms.

A “cell proliferative disorder of the lung” is a cell proliferativedisorder involving cells of the lung. Cell proliferative disorders ofthe lung can include all forms of cell proliferative disorders affectinglung cells. Cell proliferative disorders of the lung can include lungcancer, a precancer or precancerous condition of the lung, benigngrowths or lesions of the lung, and malignant growths or lesions of thelung, and metastatic lesions in tissue and organs in the body other thanthe lung. Preferably, compositions of the present invention may be usedto treat lung cancer or cell proliferative disorders of the lung. Lungcancer can include all forms of cancer of the lung. Lung cancer caninclude malignant lung neoplasms, carcinoma in situ, typical carcinoidtumors, and atypical carcinoid tumors. Lung cancer can include smallcell lung cancer (“SCLC”), non-small cell lung cancer (“NSCLC”),squamous cell carcinoma, adenocarcinoma, small cell carcinoma, largecell carcinoma, adenosquamous cell carcinoma, and mesothelioma. Lungcancer can include “scar carcinoma,” bronchioalveolar carcinoma, giantcell carcinoma, spindle cell carcinoma, and large cell neuroendocrinecarcinoma. Lung cancer can include lung neoplasms having histologic andultrastructual heterogeneity (e.g., mixed cell types).

Cell proliferative disorders of the lung can include all forms of cellproliferative disorders affecting lung cells. Cell proliferativedisorders of the lung can include lung cancer, precancerous conditionsof the lung. Cell proliferative disorders of the lung can includehyperplasia, metaplasia, and dysplasia of the lung. Cell proliferativedisorders of the lung can include asbestos-induced hyperplasia, squamousmetaplasia, and benign reactive mesothelial metaplasia. Cellproliferative disorders of the lung can include replacement of columnarepithelium with stratified squamous epithelium, and mucosal dysplasia.Individuals exposed to inhaled injurious environmental agents such ascigarette smoke and asbestos may be at increased risk for developingcell proliferative disorders of the lung. Prior lung diseases that maypredispose individuals to development of cell proliferative disorders ofthe lung can include chronic interstitial lung disease, necrotizingpulmonary disease, scleroderma, rheumatoid disease, sarcoidosis,interstitial pneumonitis, tuberculosis, repeated pneumonias, idiopathicpulmonary fibrosis, granulomata, asbestosis, fibrosing alveolitis, andHodgkin's disease.

A “cell proliferative disorder of the colon” is a cell proliferativedisorder involving cells of the colon. Preferably, the cellproliferative disorder of the colon is colon cancer. Preferably,compositions of the present invention may be used to treat colon canceror cell proliferative disorders of the colon. Colon cancer can includeall forms of cancer of the colon. Colon cancer can include sporadic andhereditary colon cancers. Colon cancer can include malignant colonneoplasms, carcinoma in situ, typical carcinoid tumors, and atypicalcarcinoid tumors. Colon cancer can include adenocarcinoma, squamous cellcarcinoma, and adenosquamous cell carcinoma. Colon cancer can beassociated with a hereditary syndrome selected from the group consistingof hereditary nonpolyposis colorectal cancer, familial adenomatouspolyposis, Gardner's syndrome, Peutz-Jeghers syndrome, Turcot's syndromeand juvenile polyposis. Colon cancer can be caused by a hereditarysyndrome selected from the group consisting of hereditary nonpolyposiscolorectal cancer, familial adenomatous polyposis, Gardner's syndrome,Peutz-Jeghers syndrome, Turcot's syndrome and juvenile polyposis.

Cell proliferative disorders of the colon can include all forms of cellproliferative disorders affecting colon cells. Cell proliferativedisorders of the colon can include colon cancer, precancerous conditionsof the colon, adenomatous polyps of the colon and metachronous lesionsof the colon. A cell proliferative disorder of the colon can includeadenoma. Cell proliferative disorders of the colon can be characterizedby hyperplasia, metaplasia, and dysplasia of the colon. Prior colondiseases that may predispose individuals to development of cellproliferative disorders of the colon can include prior colon cancer.Current disease that may predispose individuals to development of cellproliferative disorders of the colon can include Crohn's disease andulcerative colitis. A cell proliferative disorder of the colon can beassociated with a mutation in a gene selected from the group consistingof p53, ras, FAP and DCC. An individual can have an elevated risk ofdeveloping a cell proliferative disorder of the colon due to thepresence of a mutation in a gene selected from the group consisting ofp53, ras, FAP and DCC.

A “cell proliferative disorder of the pancreas” is a cell proliferativedisorder involving cells of the pancreas. Cell proliferative disordersof the pancreas can include all forms of cell proliferative disordersaffecting pancreatic cells. Cell proliferative disorders of the pancreascan include pancreas cancer, a precancer or precancerous condition ofthe pancreas, hyperplasia of the pancreas, and dysaplasia of thepancreas, benign growths or lesions of the pancreas, and malignantgrowths or lesions of the pancreas, and metastatic lesions in tissue andorgans in the body other than the pancreas. Pancreatic cancer includesall forms of cancer of the pancreas. Pancreatic cancer can includeductal adenocarcinoma, adenosquamous carcinoma, pleomorphic giant cellcarcinoma, mucinous adenocarcinoma, osteoclast-like giant cellcarcinoma, mucinous cystadenocarcinoma, acinar carcinoma, unclassifiedlarge cell carcinoma, small cell carcinoma, pancreatoblastoma, papillaryneoplasm, mucinous cystadenoma, papillary cystic neoplasm, and serouscystadenoma. Pancreatic cancer can also include pancreatic neoplasmshaving histologic and ultrastructual heterogeneity (e.g., mixed celltypes).

A “cell proliferative disorder of the prostate” is a cell proliferativedisorder involving cells of the prostate. Cell proliferative disordersof the prostate can include all forms of cell proliferative disordersaffecting prostate cells. Cell proliferative disorders of the prostatecan include prostate cancer, a precancer or precancerous condition ofthe prostate, benign growths or lesions of the prostate, and malignantgrowths or lesions of the prostate, and metastatic lesions in tissue andorgans in the body other than the prostate. Cell proliferative disordersof the prostate can include hyperplasia, metaplasia, and dysplasia ofthe prostate.

A “cell proliferative disorder of the skin” is a cell proliferativedisorder involving cells of the skin. Cell proliferative disorders ofthe skin can include all forms of cell proliferative disorders affectingskin cells. Cell proliferative disorders of the skin can include aprecancer or precancerous condition of the skin, benign growths orlesions of the skin, melanoma, malignant melanoma and other malignantgrowths or lesions of the skin, and metastatic lesions in tissue andorgans in the body other than the skin. Cell proliferative disorders ofthe skin can include hyperplasia, metaplasia, and dysplasia of the skin.

A “cell proliferative disorder of the ovary” is a cell proliferativedisorder involving cells of the ovary. Cell proliferative disorders ofthe ovary can include all forms of cell proliferative disordersaffecting cells of the ovary. Cell proliferative disorders of the ovarycan include a precancer or precancerous condition of the ovary, benigngrowths or lesions of the ovary, ovarian cancer, malignant growths orlesions of the ovary, and metastatic lesions in tissue and organs in thebody other than the ovary. Cell proliferative disorders of the skin caninclude hyperplasia, metaplasia, and dysplasia of cells of the ovary.

A “cell proliferative disorder of the breast” is a cell proliferativedisorder involving cells of the breast. Cell proliferative disorders ofthe breast can include all forms of cell proliferative disordersaffecting breast cells. Cell proliferative disorders of the breast caninclude breast cancer, a precancer or precancerous condition of thebreast, benign growths or lesions of the breast, and malignant growthsor lesions of the breast, and metastatic lesions in tissue and organs inthe body other than the breast. Cell proliferative disorders of thebreast can include hyperplasia, metaplasia, and dysplasia of the breast.

A cell proliferative disorder of the breast can be a precancerouscondition of the breast. Compositions of the present invention may beused to treat a precancerous condition of the breast. A precancerouscondition of the breast can include atypical hyperplasia of the breast,ductal carcinoma in situ (DCIS), intraductal carcinoma, lobularcarcinoma in situ (LCIS), lobular neoplasia, and stage 0 or grade 0growth or lesion of the breast (e.g., stage 0 or grade 0 breast cancer,or carcinoma in situ). A precancerous condition of the breast can bestaged according to the TNM classification scheme as accepted by theAmerican Joint Committee on Cancer (AJCC), where the primary tumor (T)has been assigned a stage of T0 or Tis; and where the regional lymphnodes (N) have been assigned a stage of N0; and where distant metastasis(M) has been assigned a stage of M0.

The cell proliferative disorder of the breast can be breast cancer.Preferably, compositions of the present invention may be used to treatbreast cancer. Breast cancer includes all forms of cancer of the breast.Breast cancer can include primary epithelial breast cancers. Breastcancer can include cancers in which the breast is involved by othertumors such as lymphoma, sarcoma or melanoma. Breast cancer can includecarcinoma of the breast, ductal carcinoma of the breast, lobularcarcinoma of the breast, undifferentiated carcinoma of the breast,cystosarcoma phyllodes of the breast, angiosarcoma of the breast, andprimary lymphoma of the breast. Breast cancer can include Stage I, II,IIIA, IIIB, IIIC and IV breast cancer. Ductal carcinoma of the breastcan include invasive carcinoma, invasive carcinoma in situ withpredominant intraductal component, inflammatory breast cancer, and aductal carcinoma of the breast with a histologic type selected from thegroup consisting of comedo, mucinous (colloid), medullary, medullarywith lymphcytic infiltrate, papillary, scirrhous, and tubular. Lobularcarcinoma of the breast can include invasive lobular carcinoma withpredominant in situ component, invasive lobular carcinoma, andinfiltrating lobular carcinoma. Breast cancer can include Paget'sdisease, Paget's disease with intraductal carcinoma, and Paget's diseasewith invasive ductal carcinoma. Breast cancer can include breastneoplasms having histologic and ultrastructual heterogeneity (e.g.,mixed cell types).

Preferably, compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, polymorph, or solvate thereof, maybe used to treat breast cancer. A breast cancer that is to be treatedcan include familial breast cancer. A breast cancer that is to betreated can include sporadic breast cancer. A breast cancer that is tobe treated can arise in a male subject. A breast cancer that is to betreated can arise in a female subject. A breast cancer that is to betreated can arise in a premenopausal female subject or a postmenopausalfemale subject. A breast cancer that is to be treated can arise in asubject equal to or older than 30 years old, or a subject younger than30 years old. A breast cancer that is to be treated has arisen in asubject equal to or older than 50 years old, or a subject younger than50 years old. A breast cancer that is to be treated can arise in asubject equal to or older than 70 years old, or a subject younger than70 years old.

A breast cancer that is to be treated can be typed to identify afamilial or spontaneous mutation in BRCA1, BRCA2, or p53. A breastcancer that is to be treated can be typed as having a HER2/neu geneamplification, as overexpressing HER2/neu, or as having a low,intermediate or high level of HER2/neu expression. A breast cancer thatis to be treated can be typed for a marker selected from the groupconsisting of estrogen receptor (ER), progesterone receptor (PR), humanepidermal growth factor receptor-2, Ki-67, CA15-3, CA 27-29, and c-Met.A breast cancer that is to be treated can be typed as ER-unknown,ER-rich or ER-poor. A breast cancer that is to be treated can be typedas ER-negative or ER-positive. ER-typing of a breast cancer may beperformed by any reproducible means. ER-typing of a breast cancer may beperformed as set forth in Onkologie 27: 175-179 (2004). A breast cancerthat is to be treated can be typed as PR-unknown, PR-rich, or PR-poor. Abreast cancer that is to be treated can be typed as PR-negative orPR-positive. A breast cancer that is to be treated can be typed asreceptor positive or receptor negative. A breast cancer that is to betreated can be typed as being associated with elevated blood levels ofCA 15-3, or CA 27-29, or both.

A breast cancer that is to be treated can include a localized tumor ofthe breast. A breast cancer that is to be treated can include a tumor ofthe breast that is associated with a negative sentinel lymph node (SLN)biopsy. A breast cancer that is to be treated can include a tumor of thebreast that is associated with a positive sentinel lymph node (SLN)biopsy. A breast cancer that is to be treated can include a tumor of thebreast that is associated with one or more positive axillary lymphnodes, where the axillary lymph nodes have been staged by any applicablemethod. A breast cancer that is to be treated can include a tumor of thebreast that has been typed as having nodal negative status (e.g.,node-negative) or nodal positive status (e.g., node-positive). A breastcancer that is to be treated can include a tumor of the breast that hasmetastasized to other locations in the body. A breast cancer that is tobe treated can be classified as having metastasized to a locationselected from the group consisting of bone, lung, liver, or brain. Abreast cancer that is to be treated can be classified according to acharacteristic selected from the group consisting of metastatic,localized, regional, local-regional, locally advanced, distant,multicentric, bilateral, ipsilateral, contralateral, newly diagnosed,recurrent, and inoperable.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, may be used totreat or prevent a cell proliferative disorder of the breast, or totreat or prevent breast cancer, in a subject having an increased risk ofdeveloping breast cancer relative to the population at large. A subjectwith an increased risk of developing breast cancer relative to thepopulation at large is a female subject with a family history orpersonal history of breast cancer. A subject with an increased risk ofdeveloping breast cancer relative to the population at large is a femalesubject having a germ-line or spontaneous mutation in BRCA1 or BRCA2, orboth. A subject with an increased risk of developing breast cancerrelative to the population at large is a female subject with a familyhistory of breast cancer and a germ-line or spontaneous mutation inBRCA1 or BRCA2, or both. A subject with an increased risk of developingbreast cancer relative to the population at large is a female who isgreater than 30 years old, greater than 40 years old, greater than 50years old, greater than 60 years old, greater than 70 years old, greaterthan 80 years old, or greater than 90 years old. A subject with anincreased risk of developing breast cancer relative to the population atlarge is a subject with atypical hyperplasia of the breast, ductalcarcinoma in situ (DCIS), intraductal carcinoma, lobular carcinoma insitu (LCIS), lobular neoplasia, or a stage 0 growth or lesion of thebreast (e.g., stage 0 or grade 0 breast cancer, or carcinoma in situ).

A breast cancer that is to be treated can histologically gradedaccording to the Scarff-Bloom-Richardson system, wherein a breast tumorhas been assigned a mitosis count score of 1, 2, or 3; a nuclearpleiomorphism score of 1, 2, or 3; a tubule formation score of 1, 2, or3; and a total Scarff-Bloom-Richardson score of between 3 and 9. Abreast cancer that is to be treated can be assigned a tumor gradeaccording to the International Consensus Panel on the Treatment ofBreast Cancer selected from the group consisting of grade 1, grade 1-2,grade 2, grade 2-3, or grade 3.

A cancer that is to be treated can be staged according to the AmericanJoint Committee on Cancer (AJCC) TNM classification system, where thetumor (T) has been assigned a stage of TX, T1, T1mic, T1a, T1b, T1c, T2,T3, T4, T4a, T4b, T4c, or T4d; and where the regional lymph nodes (N)have been assigned a stage of NX, N0, N1, N2, N2a, N2b, N3, N3a, N3b, orN3c; and where distant metastasis (M) can be assigned a stage of MX, M0,or M1. A cancer that is to be treated can be staged according to anAmerican Joint Committee on Cancer (AJCC) classification as Stage I,Stage IIA, Stage IIB, Stage IIIA, Stage IIIB, Stage IIIC, or Stage IV. Acancer that is to be treated can be assigned a grade according to anAJCC classification as Grade GX (e.g., grade cannot be assessed), Grade1, Grade 2, Grade 3 or Grade 4. A cancer that is to be treated can bestaged according to an AJCC pathologic classification (pN) of pNX, pN0,PN0 (I−), PN0 (I+), PN0 (mol−), PN0 (mol+), PN1, PN1(mi), PN1a, PN1b,PN1c, pN2, pN2a, pN2b, pN3, pN3a, pN3b, or pN3c.

A cancer that is to be treated can include a tumor that has beendetermined to be less than or equal to about 2 centimeters in diameter.A cancer that is to be treated can include a tumor that has beendetermined to be from about 2 to about 5 centimeters in diameter. Acancer that is to be treated can include a tumor that has beendetermined to be greater than or equal to about 3 centimeters indiameter. A cancer that is to be treated can include a tumor that hasbeen determined to be greater than 5 centimeters in diameter. A cancerthat is to be treated can be classified by microscopic appearance aswell differentiated, moderately differentiated, poorly differentiated,or undifferentiated. A cancer that is to be treated can be classified bymicroscopic appearance with respect to mitosis count (e.g., amount ofcell division) or nuclear pleiomorphism (e.g., change in cells). Acancer that is to be treated can be classified by microscopic appearanceas being associated with areas of necrosis (e.g., areas of dying ordegenerating cells). A cancer that is to be treated can be classified ashaving an abnormal karyotype, having an abnormal number of chromosomes,or having one or more chromosomes that are abnormal in appearance. Acancer that is to be treated can be classified as being aneuploid,triploid, tetraploid, or as having an altered ploidy. A cancer that isto be treated can be classified as having a chromosomal translocation,or a deletion or duplication of an entire chromosome, or a region ofdeletion, duplication or amplification of a portion of a chromosome.

A cancer that is to be treated can be evaluated by DNA cytometry, flowcytometry, or image cytometry. A cancer that is to be treated can betyped as having 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of cellsin the synthesis stage of cell division (e.g., in S phase of celldivision). A cancer that is to be treated can be typed as having a lowS-phase fraction or a high S-phase fraction.

As used herein, a “normal cell” is a cell that cannot be classified aspart of a “cell proliferative disorder”. A normal cell lacks unregulatedor abnormal growth, or both, that can lead to the development of anunwanted condition or disease. Preferably, a normal cell possessesnormally functioning cell cycle checkpoint control mechanisms.

As used herein, “contacting a cell” refers to a condition in which acompound or other composition of matter is in direct contact with acell, or is close enough to induce a desired biological effect in acell.

As used herein, “candidate compound” refers to a compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,polymorph or solvate thereof, that has been or will be tested in one ormore in vitro or in vivo biological assays, in order to determine ifthat compound is likely to elicit a desired biological or medicalresponse in a cell, tissue, system, animal or human that is being soughtby a researcher or clinician. A candidate compound is a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof. The biological or medicalresponse can be the treatment of cancer. The biological or medicalresponse can be treatment or prevention of a cell proliferativedisorder. In vitro or in vivo biological assays can include, but are notlimited to, enzymatic activity assays, electrophoretic mobility shiftassays, reporter gene assays, in vitro cell viability assays, and theassays described herein.

As used herein, “monotherapy” refers to the administration of a singleactive or therapeutic compound to a subject in need thereof. Preferably,monotherapy will involve administration of a therapeutically effectiveamount of an active compound. For example, cancer monotherapy with oneof the compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, analog or derivative thereof, to asubject in need of treatment of cancer. Monotherapy may be contrastedwith combination therapy, in which a combination of multiple activecompounds is administered, preferably with each component of thecombination present in a therapeutically effective amount. In oneaspect, monotherapy with a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, is more effective than combination therapy in inducinga desired biological effect.

As used herein, “treating” or “treat” describes the management and careof a patient for the purpose of combating a disease, condition, ordisorder and includes the administration of a compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,polymorph or solvate thereof, to alleviate the symptoms or complicationsof a disease, condition or disorder, or to eliminate the disease,condition or disorder.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, can also beused to prevent a disease, condition or disorder. As used herein,“preventing” or “prevent” describes reducing or eliminating the onset ofthe symptoms or complications of the disease, condition or disorder.

As used herein, the term “alleviate” is meant to describe a process bywhich the severity of a sign or symptom of a disorder is decreased.Importantly, a sign or symptom can be alleviated without beingeliminated. In a preferred embodiment, the administration ofpharmaceutical compositions of the invention leads to the elimination ofa sign or symptom, however, elimination is not required. Effectivedosages are expected to decrease the severity of a sign or symptom. Forinstance, a sign or symptom of a disorder such as cancer, which canoccur in multiple locations, is alleviated if the severity of the canceris decreased within at least one of multiple locations.

As used herein, the term “severity” is meant to describe the potentialof cancer to transform from a precancerous, or benign, state into amalignant state. Alternatively, or in addition, severity is meant todescribe a cancer stage, for example, according to the TNM system(accepted by the International Union Against Cancer (UICC) and theAmerican Joint Committee on Cancer (AJCC)) or by other art-recognizedmethods. Cancer stage refers to the extent or severity of the cancer,based on factors such as the location of the primary tumor, tumor size,number of tumors, and lymph node involvement (spread of cancer intolymph nodes). Alternatively, or in addition, severity is meant todescribe the tumor grade by art-recognized methods (see, National CancerInstitute, www.cancer.gov). Tumor grade is a system used to classifycancer cells in terms of how abnormal they look under a microscope andhow quickly the tumor is likely to grow and spread. Many factors areconsidered when determining tumor grade, including the structure andgrowth pattern of the cells. The specific factors used to determinetumor grade vary with each type of cancer. Severity also describes ahistologic grade, also called differentiation, which refers to how muchthe tumor cells resemble normal cells of the same tissue type (see,National Cancer Institute, www.cancer.gov). Furthermore, severitydescribes a nuclear grade, which refers to the size and shape of thenucleus in tumor cells and the percentage of tumor cells that aredividing (see, National Cancer Institute, www.cancer.gov).

In another aspect of the invention, severity describes the degree towhich a tumor has secreted growth factors, degraded the extracellularmatrix, become vascularized, lost adhesion to juxtaposed tissues, ormetastasized. Moreover, severity describes the number of locations towhich a primary tumor has metastasized. Finally, severity includes thedifficulty of treating tumors of varying types and locations. Forexample, inoperable tumors, those cancers which have greater access tomultiple body systems (hematological and immunological tumors), andthose which are the most resistant to traditional treatments areconsidered most severe. In these situations, prolonging the lifeexpectancy of the subject and/or reducing pain, decreasing theproportion of cancerous cells or restricting cells to one system, andimproving cancer stage/tumor grade/histological grade/nuclear grade areconsidered alleviating a sign or symptom of the cancer.

As used herein the term “symptom” is defined as an indication ofdisease, illness, injury, or that something is not right in the body.Symptoms are felt or noticed by the individual experiencing the symptom,but may not easily be noticed by others. Others are defined asnon-health-care professionals.

As used herein the term “sign” is also defined as an indication thatsomething is not right in the body. But signs are defined as things thatcan be seen by a doctor, nurse, or other health care professional.

Cancer is a group of diseases that may cause almost any sign or symptom.The signs and symptoms will depend on where the cancer is, the size ofthe cancer, and how much it affects the nearby organs or structures. Ifa cancer spreads (metastasizes), then symptoms may appear in differentparts of the body.

As a cancer grows, it begins to push on nearby organs, blood vessels,and nerves. This pressure creates some of the signs and symptoms ofcancer. If the cancer is in a critical area, such as certain parts ofthe brain, even the smallest tumor can cause early symptoms.

But sometimes cancers start in places where it does not cause anysymptoms until the cancer has grown quite large. Pancreas cancers, forexample, do not usually grow large enough to be felt from the outside ofthe body. Some pancreatic cancers do not cause symptoms until they beginto grow around nearby nerves (this causes a backache). Others growaround the bile duct, which blocks the flow of bile and leads to ayellowing of the skin known as jaundice. By the time a pancreatic cancercauses these signs or symptoms, it has usually reached an advancedstage.

A cancer may also cause symptoms such as fever, fatigue, or weight loss.This may be because cancer cells use up much of the body's energy supplyor release substances that change the body's metabolism. Or the cancermay cause the immune system to react in ways that produce thesesymptoms.

Sometimes, cancer cells release substances into the bloodstream thatcause symptoms not usually thought to result from cancers. For example,some cancers of the pancreas can release substances which cause bloodclots to develop in veins of the legs. Some lung cancers makehormone-like substances that affect blood calcium levels, affectingnerves and muscles and causing weakness and dizziness

Cancer presents several general signs or symptoms that occur when avariety of subtypes of cancer cells are present. Most people with cancerwill lose weight at some time with their disease. An unexplained(unintentional) weight loss of 10 pounds or more may be the first signof cancer, particularly cancers of the pancreas, stomach, esophagus, orlung.

Fever is very common with cancer, but is more often seen in advanceddisease. Almost all patients with cancer will have fever at some time,especially if the cancer or its treatment affects the immune system andmakes it harder for the body to fight infection. Less often, fever maybe an early sign of cancer, such as with leukemia or lymphoma.

Fatigue may be an important symptom as cancer progresses. It may happenearly, though, in cancers such as with leukemia, or if the cancer iscausing an ongoing loss of blood, as in some colon or stomach cancers.

Pain may be an early symptom with some cancers such as bone cancers ortesticular cancer. But most often pain is a symptom of advanced disease.

Along with cancers of the skin (see next section), some internal cancerscan cause skin signs that can be seen. These changes include the skinlooking darker (hyperpigmentation), yellow (jaundice), or red(erythema); itching; or excessive hair growth.

Alternatively, or in addition, cancer subtypes present specific signs orsymptoms. Changes in bowel habits or bladder function could indicatecancer. Long-term constipation, diarrhea, or a change in the size of thestool may be a sign of colon cancer. Pain with urination, blood in theurine, or a change in bladder function (such as more frequent or lessfrequent urination) could be related to bladder or prostate cancer.

Changes in skin condition or appearance of a new skin condition couldindicate cancer. Skin cancers may bleed and look like sores that do notheal. A long-lasting sore in the mouth could be an oral cancer,especially in patients who smoke, chew tobacco, or frequently drinkalcohol. Sores on the penis or vagina may either be signs of infectionor an early cancer.

Unusual bleeding or discharge could indicate cancer. Unusual bleedingcan happen in either early or advanced cancer. Blood in the sputum(phlegm) may be a sign of lung cancer. Blood in the stool (or a dark orblack stool) could be a sign of colon or rectal cancer. Cancer of thecervix or the endometrium (lining of the uterus) can cause vaginalbleeding. Blood in the urine may be a sign of bladder or kidney cancer.A bloody discharge from the nipple may be a sign of breast cancer.

A thickening or lump in the breast or in other parts of the body couldindicate the presence of a cancer. Many cancers can be felt through theskin, mostly in the breast, testicle, lymph nodes (glands), and the softtissues of the body. A lump or thickening may be an early or late signof cancer. Any lump or thickening could be indicative of cancer,especially if the formation is new or has grown in size.

Indigestion or trouble swallowing could indicate cancer. While thesesymptoms commonly have other causes, indigestion or swallowing problemsmay be a sign of cancer of the esophagus, stomach, or pharynx (throat).

Recent changes in a wart or mole could be indicative of cancer. Anywart, mole, or freckle that changes in color, size, or shape, or losesits definite borders indicates the potential development of cancer. Forexample, the skin lesion may be a melanoma.

A persistent cough or hoarseness could be indicative of cancer. A coughthat does not go away may be a sign of lung cancer. Hoarseness can be asign of cancer of the larynx (voice box) or thyroid.

While the signs and symptoms listed above are the more common ones seenwith cancer, there are many others that are less common and are notlisted here. However, all art-recognized signs and symptoms of cancerare contemplated and encompassed by the instant invention.

Treating cancer can result in a reduction in size of a tumor. Areduction in size of a tumor may also be referred to as “tumorregression”. Preferably, after treatment, tumor size is reduced by 5% orgreater relative to its size prior to treatment; more preferably, tumorsize is reduced by 10% or greater; more preferably, reduced by 20% orgreater; more preferably, reduced by 30% or greater; more preferably,reduced by 40% or greater; even more preferably, reduced by 50% orgreater; and most preferably, reduced by greater than 75% or greater.Size of a tumor may be measured by any reproducible means ofmeasurement. The size of a tumor may be measured as a diameter of thetumor.

Treating cancer can result in a reduction in tumor volume. Preferably,after treatment, tumor volume is reduced by 5% or greater relative toits size prior to treatment; more preferably, tumor volume is reduced by10% or greater; more preferably, reduced by 20% or greater; morepreferably, reduced by 30% or greater; more preferably, reduced by 40%or greater; even more preferably, reduced by 50% or greater; and mostpreferably, reduced by greater than 75% or greater. Tumor volume may bemeasured by any reproducible means of measurement.

Treating cancer results in a decrease in number of tumors. Preferably,after treatment, tumor number is reduced by 5% or greater relative tonumber prior to treatment; more preferably, tumor number is reduced by10% or greater; more preferably, reduced by 20% or greater; morepreferably, reduced by 30% or greater; more preferably, reduced by 40%or greater; even more preferably, reduced by 50% or greater; and mostpreferably, reduced by greater than 75%. Number of tumors may bemeasured by any reproducible means of measurement. The number of tumorsmay be measured by counting tumors visible to the naked eye or at aspecified magnification. Preferably, the specified magnification is 2×,3×, 4×, 5×, 10×, or 50×.

Treating cancer can result in a decrease in number of metastatic lesionsin other tissues or organs distant from the primary tumor site.Preferably, after treatment, the number of metastatic lesions is reducedby 5% or greater relative to number prior to treatment; more preferably,the number of metastatic lesions is reduced by 10% or greater; morepreferably, reduced by 20% or greater; more preferably, reduced by 30%or greater; more preferably, reduced by 40% or greater; even morepreferably, reduced by 50% or greater; and most preferably, reduced bygreater than 75%. The number of metastatic lesions may be measured byany reproducible means of measurement. The number of metastatic lesionsmay be measured by counting metastatic lesions visible to the naked eyeor at a specified magnification. Preferably, the specified magnificationis 2×, 3×, 4×, 5×, 10×, or 50×.

Treating cancer can result in an increase in average survival time of apopulation of treated subjects in comparison to a population receivingcarrier alone. Preferably, the average survival time is increased bymore than 30 days; more preferably, by more than 60 days; morepreferably, by more than 90 days; and most preferably, by more than 120days. An increase in average survival time of a population may bemeasured by any reproducible means. An increase in average survival timeof a population may be measured, for example, by calculating for apopulation the average length of survival following initiation oftreatment with an active compound. An increase in average survival timeof a population may also be measured, for example, by calculating for apopulation the average length of survival following completion of afirst round of treatment with an active compound.

Treating cancer can result in an increase in average survival time of apopulation of treated subjects in comparison to a population ofuntreated subjects. Preferably, the average survival time is increasedby more than 30 days; more preferably, by more than 60 days; morepreferably, by more than 90 days; and most preferably, by more than 120days. An increase in average survival time of a population may bemeasured by any reproducible means. An increase in average survival timeof a population may be measured, for example, by calculating for apopulation the average length of survival following initiation oftreatment with an active compound. An increase in average survival timeof a population may also be measured, for example, by calculating for apopulation the average length of survival following completion of afirst round of treatment with an active compound.

Treating cancer can result in increase in average survival time of apopulation of treated subjects in comparison to a population receivingmonotherapy with a drug that is not a compound of the present invention,or a pharmaceutically acceptable salt, prodrug, metabolite, analog orderivative thereof. Preferably, the average survival time is increasedby more than 30 days; more preferably, by more than 60 days; morepreferably, by more than 90 days; and most preferably, by more than 120days. An increase in average survival time of a population may bemeasured by any reproducible means. An increase in average survival timeof a population may be measured, for example, by calculating for apopulation the average length of survival following initiation oftreatment with an active compound. An increase in average survival timeof a population may also be measured, for example, by calculating for apopulation the average length of survival following completion of afirst round of treatment with an active compound.

Treating cancer can result in a decrease in the mortality rate of apopulation of treated subjects in comparison to a population receivingcarrier alone. Treating cancer can result in a decrease in the mortalityrate of a population of treated subjects in comparison to an untreatedpopulation. Treating cancer can result in a decrease in the mortalityrate of a population of treated subjects in comparison to a populationreceiving monotherapy with a drug that is not a compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,analog or derivative thereof. Preferably, the mortality rate isdecreased by more than 2%; more preferably, by more than 5%; morepreferably, by more than 10%; and most preferably, by more than 25%. Adecrease in the mortality rate of a population of treated subjects maybe measured by any reproducible means. A decrease in the mortality rateof a population may be measured, for example, by calculating for apopulation the average number of disease-related deaths per unit timefollowing initiation of treatment with an active compound. A decrease inthe mortality rate of a population may also be measured, for example, bycalculating for a population the average number of disease-relateddeaths per unit time following completion of a first round of treatmentwith an active compound.

Treating cancer can result in a decrease in tumor growth rate.Preferably, after treatment, tumor growth rate is reduced by at least 5%relative to number prior to treatment; more preferably, tumor growthrate is reduced by at least 10%; more preferably, reduced by at least20%; more preferably, reduced by at least 30%; more preferably, reducedby at least 40%; more preferably, reduced by at least 50%; even morepreferably, reduced by at least 50%; and most preferably, reduced by atleast 75%. Tumor growth rate may be measured by any reproducible meansof measurement. Tumor growth rate can be measured according to a changein tumor diameter per unit time.

Treating cancer can result in a decrease in tumor regrowth. Preferably,after treatment, tumor regrowth is less than 5%; more preferably, tumorregrowth is less than 10%; more preferably, less than 20%; morepreferably, less than 30%; more preferably, less than 40%; morepreferably, less than 50%; even more preferably, less than 50%; and mostpreferably, less than 75%. Tumor regrowth may be measured by anyreproducible means of measurement. Tumor regrowth is measured, forexample, by measuring an increase in the diameter of a tumor after aprior tumor shrinkage that followed treatment. A decrease in tumorregrowth is indicated by failure of tumors to reoccur after treatmenthas stopped.

Treating or preventing a cell proliferative disorder can result in areduction in the rate of cellular proliferation. Preferably, aftertreatment, the rate of cellular proliferation is reduced by at least 5%;more preferably, by at least 10%; more preferably, by at least 20%; morepreferably, by at least 30%; more preferably, by at least 40%; morepreferably, by at least 50%; even more preferably, by at least 50%; andmost preferably, by at least 75%. The rate of cellular proliferation maybe measured by any reproducible means of measurement. The rate ofcellular proliferation is measured, for example, by measuring the numberof dividing cells in a tissue sample per unit time.

Treating or preventing a cell proliferative disorder can result in areduction in the proportion of proliferating cells. Preferably, aftertreatment, the proportion of proliferating cells is reduced by at least5%; more preferably, by at least 10%; more preferably, by at least 20%;more preferably, by at least 30%; more preferably, by at least 40%; morepreferably, by at least 50%; even more preferably, by at least 50%; andmost preferably, by at least 75%. The proportion of proliferating cellsmay be measured by any reproducible means of measurement. Preferably,the proportion of proliferating cells is measured, for example, byquantifying the number of dividing cells relative to the number ofnondividing cells in a tissue sample. The proportion of proliferatingcells can be equivalent to the mitotic index.

Treating or preventing a cell proliferative disorder can result in adecrease in size of an area or zone of cellular proliferation.Preferably, after treatment, size of an area or zone of cellularproliferation is reduced by at least 5% relative to its size prior totreatment; more preferably, reduced by at least 10%; more preferably,reduced by at least 20%; more preferably, reduced by at least 30%; morepreferably, reduced by at least 40%; more preferably, reduced by atleast 50%; even more preferably, reduced by at least 50%; and mostpreferably, reduced by at least 75%. Size of an area or zone of cellularproliferation may be measured by any reproducible means of measurement.The size of an area or zone of cellular proliferation may be measured asa diameter or width of an area or zone of cellular proliferation.

Treating or preventing a cell proliferative disorder can result in adecrease in the number or proportion of cells having an abnormalappearance or morphology. Preferably, after treatment, the number ofcells having an abnormal morphology is reduced by at least 5% relativeto its size prior to treatment; more preferably, reduced by at least10%; more preferably, reduced by at least 20%; more preferably, reducedby at least 30%; more preferably, reduced by at least 40%; morepreferably, reduced by at least 50%; even more preferably, reduced by atleast 50%; and most preferably, reduced by at least 75%. An abnormalcellular appearance or morphology may be measured by any reproduciblemeans of measurement. An abnormal cellular morphology can be measured bymicroscopy, e.g., using an inverted tissue culture microscope. Anabnormal cellular morphology can take the form of nuclear pleiomorphism.

As used herein, the term “selectively” means tending to occur at ahigher frequency in one population than in another population. Thecompared populations can be cell populations. Preferably, a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, acts selectively on a canceror precancerous cell but not on a normal cell. Preferably, a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, acts selectively to modulateone molecular target (e.g., a target protein methyltransferase) but doesnot significantly modulate another molecular target (e.g., a non-targetprotein methyltransferase). The invention also provides a method forselectively inhibiting the activity of an enzyme, such as a proteinmethyltransferase. Preferably, an event occurs selectively in populationA relative to population B if it occurs greater than two times morefrequently in population A as compared to population B. An event occursselectively if it occurs greater than five times more frequently inpopulation A. An event occurs selectively if it occurs greater than tentimes more frequently in population A; more preferably, greater thanfifty times; even more preferably, greater than 100 times; and mostpreferably, greater than 1000 times more frequently in population A ascompared to population B. For example, cell death would be said to occurselectively in cancer cells if it occurred greater than twice asfrequently in cancer cells as compared to normal cells.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, can modulatethe activity of a molecular target (e.g., a target proteinmethyltransferase). Modulating refers to stimulating or inhibiting anactivity of a molecular target. Preferably, a compound of the presentinvention, or a pharmaceutically acceptable salt, prodrug, metabolite,polymorph or solvate thereof, modulates the activity of a moleculartarget if it stimulates or inhibits the activity of the molecular targetby at least 2-fold relative to the activity of the molecular targetunder the same conditions but lacking only the presence of saidcompound. More preferably, a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, modulates the activity of a molecular target if itstimulates or inhibits the activity of the molecular target by at least5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least100-fold relative to the activity of the molecular target under the sameconditions but lacking only the presence of said compound. The activityof a molecular target may be measured by any reproducible means. Theactivity of a molecular target may be measured in vitro or in vivo. Forexample, the activity of a molecular target may be measured in vitro byan enzymatic activity assay or a DNA binding assay, or the activity of amolecular target may be measured in vivo by assaying for expression of areporter gene.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, does notsignificantly modulate the activity of a molecular target if theaddition of the compound does not stimulate or inhibit the activity ofthe molecular target by greater than 10% relative to the activity of themolecular target under the same conditions but lacking only the presenceof said compound.

As used herein, the term “isozyme selective” means preferentialinhibition or stimulation of a first isoform of an enzyme in comparisonto a second isoform of an enzyme (e.g., preferential inhibition orstimulation of a protein methyltransferase isozyme alpha in comparisonto a protein methyltransferase isozyme beta). Preferably, a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, demonstrates a minimum of afourfold differential, preferably a tenfold differential, morepreferably a fifty fold differential, in the dosage required to achievea biological effect. Preferably, a compound of the present invention, ora pharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, demonstrates this differential across the range ofinhibition, and the differential is exemplified at the IC₅₀, i.e., a 50%inhibition, for a molecular target of interest.

Administering a compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, polymorph or solvate thereof, to acell or a subject in need thereof can result in modulation (i.e.,stimulation or inhibition) of an activity of a protein methyltransferaseof interest.

The present invention provides methods to assess biological activity ofa compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof or methods ofidentifying a test compound as an inhibitor of a Y641 mutant of EZH2. Inone embodiment the method includes combining an isolated Y641 mutant ofEZH2 with a histone substrate, a methyl group donor (such asS-adenosylmethionine (SAM)), and a test compound, wherein the histonesubstrate comprises a form of H3-K27 selected from the group consistingof unmethylated H3-K27, monomethylated H3-K27, dimethylated H3-K27, andany combination thereof; and performing an assay to detect methylationof H3-K27 in the histone substrate, thereby identifying the testcompound as an inhibitor of the Y641 mutant of EZH2 when methylation ofH3-K27 in the presence of the test compound is less than methylation ofH3-K27 in the absence of the test compound. The assay to detectmethylation of H3-K27 can be selected to measure the rate ofmethylation, the extent of methylation, or both the rate and extent ofmethylation.

The Y641 mutant of EZH2 is isolated as a PRC2 complex or functionalequivalent thereof. As used herein, the term “isolated” meanssubstantially separated from other components with which the complex maybe found as it occurs in nature. A compound can be isolated withoutnecessarily being purified. In one embodiment the mutant of EZH2 isisolated as a complex of a Y641 mutant of EZH2 together with EED andSUZ12. In another embodiment the mutant of EZH2 is isolated as a complexof a Y641 mutant of EZH2 together with EED, SUZ12, and RbAp48. Underappropriate conditions, a PRC2 complex or functional equivalent thereofexhibits histone methyltransferase activity for H3-K27. In oneembodiment the complex is composed of recombinantly expressed componentpolypeptides, e.g., EZH2, EED, SUZ12, with or without RbAp48.

The isolated Y641 mutant of EZH2 is combined with a histone substrate. Ahistone substrate includes any suitable source of histone polypeptidesor fragments thereof that can serve as substrate for EZH2. In oneembodiment the histone substrate includes histones isolated from asubject. The histones can be isolated from cells of a subject using anysuitable method; such methods are well known to persons skilled in theart and need not be further specified here. See, for example, Fang etal. (2004) Methods Enzymol 377:213-26. In accordance with the Examplesbelow, in one embodiment the histone substrate is provided asnucleosomes. In accordance with the Examples below, in one embodimentthe histone substrate is provided as avian (chicken) erythrocytenucleosomes.

Histone substrate so provided may include an admixture of states ofhistone modification, including various states of H3-K27 methylation asjudged by Western blotting with H3-K27 methylation state-specificantibodies. In one embodiment the histone substrate may be provided aspurified full-length histone H3. Such purified full-length histone H3may be provided as a homogeneous preparation in respect of states ofH3-K27 methylation, or as an admixture of various states of H3-K27methylation. Homogeneous preparations of isolated histone H3 in respectof states of H3-K27 methylation may be prepared in part by passage overan immunoaffinity column loaded with suitable H3-K27 methylationstate-specific antibodies or by immunoprecipitation using magnetic beadscoated with suitable H3-K27 methylation state-specific antibodies.Alternatively or in addition, the methylation state of H3-K27 can becharacterized as part of performing the assay. For example, the startingmaterial histone substrate might be characterized as containing 50percent unmethylated H3-K27, 40 percent monomethylated H3-K27, 10percent dimethylated H3-K27, and 0 percent trimethylated H3-K27.

In one embodiment the histone substrate includes a peptide library or asuitable peptide comprising one or more amino acid sequences related tohistone H3, including, in particular, a sequence that encompassesH3-K27. For example, in one embodiment, the histone substrate is apeptide fragment that corresponds to amino acid residues 21-44 ofhistone H3. The peptide library or peptide can be prepared by peptidesynthesis according to techniques well known in the art and optionallymodified so as to incorporate any desired degree of methylation oflysine corresponding to H3-K27. As described in the Examples below, suchpeptides can also be modified to incorporate a label, such as biotin,useful in performing downstream assays. In one embodiment the label isappended to the amino (N)-terminus of the peptide(s). In one embodimentthe label is appended to the carboxy (C)-terminus of the peptide(s).

Detection of methylation of H3-K27 can be accomplished using anysuitable method. In one embodiment, the source of donor methyl groupsincludes methyl groups that are labeled with a detectable label. Thedetectable label in one embodiment is an isotopic label, e.g., tritium.Other types of labels may include, for example, fluorescent labels.

Detection of formation of trimethylated H3-K27 can be accomplished usingany suitable method. For example, detection of formation oftrimethylated H3-K27 can be accomplished using an assay to detectincorporation of labeled methyl groups, such as described above,optionally combined with a chromatographic or other method to separatelabeled products by size, e.g., polyacrylamide gel electrophoresis(PAGE), capillary electrophoresis (CE), or high pressure liquidchromatography (HPLC). Alternatively or in addition, detection offormation of trimethylated H3-K27 can be accomplished using antibodiesthat are specific for trimethylated H3-K27.

Detection of conversion of monomethylated H3-K27 to dimethylated H3-K27can be accomplished using any suitable method. In one embodiment theconversion is measured using antibodies specific for monomethylatedH3-K27 and dimethylated H3-K27. For example, starting amounts orconcentrations of monomethylated H3-K27 and dimethylated H3-K27 may bedetermined using appropriate antibodies specific for monomethylatedH3-K27 and dimethylated H3-K27. Following the combination of enzyme,substrate, methyl group donor, and test compound, resulting amounts orconcentrations of monomethylated H3-K27 and dimethylated H3-K27 may thenbe determined using appropriate antibodies specific for monomethylatedH3-K27 and dimethylated H3-K27. The beginning and resulting amounts orconcentrations of monomethylated H3-K27 and dimethylated H3-K27 can thenbe compared. Alternatively or in addition, beginning and resultingamounts or concentrations of monomethylated H3-K27 and dimethylatedH3-K27 can then be compared to corresponding amounts of concentrationsfrom a negative control. A negative control reaction, in which no testagent is included in the assay, can be run in parallel or as ahistorical control. Results of such control reaction can optionally besubtracted from corresponding results of the experimental reaction priorto or in conjunction with making the comparison mentioned above.

Because the dimethylated form of H3-K27 may be further methylated in thesame assay, a reduction in the amount or concentration of monomethylatedH3-K27 may not appear to correspond directly to an increase indimethylated H3-K27. In this instance, it may be presumed, however, thata reduction in the amount or concentration of monomethylated H3-K27 is,by itself, reflective of conversion of monomethylated H3-K27 todimethylated H3-K27.

Detection of conversion of dimethylated H3-K27 to trimethylated H3-K27can be accomplished using any suitable method. In one embodiment theconversion is measured using antibodies specific for dimethylated H3-K27and trimethylated H3-K27. For example, starting amounts orconcentrations of dimethylated H3-K27 and trimethylated H3-K27 may bedetermined using appropriate antibodies specific for dimethylated H3-K27and trimethylated H3-K27. Following the combination of enzyme,substrate, and test compound, resulting amounts or concentrations ofdimethylated H3-K27 and trimethylated H3-K27 may then be determinedusing appropriate antibodies specific for dimethylated H3-K27 andtrimethylated H3-K27. The beginning and resulting amounts orconcentrations of dimethylated H3-K27 and trimethylated H3-K27 can thenbe compared. Alternatively or in addition, beginning and resultingamounts or concentrations of dimethylated H3-K27 and trimethylatedH3-K27 can then be compared to corresponding amounts of concentrationsfrom a negative control. A negative control reaction, in which no testagent is included in the assay, can be run in parallel or as ahistorical control. Results of such control reaction can optionally besubtracted from corresponding results of the experimental reaction priorto or in conjunction with making the comparison mentioned above.

A test agent is identified as an inhibitor of the Y641 mutant of EZH2when methylation of H3-K27 with the test compound is less thanmethylation of H3-K27 without the test compound. In one embodiment, atest agent is identified as an inhibitor of the Y641 mutant of EZH2 whenformation of trimethylated H3-K27 in the presence of the test compoundis less than formation of trimethylated H3-K27 in the absence of thetest compound.

The present invention also provides a method for identifying a selectiveinhibitor of a Y641 mutant of EZH2. In one embodiment the methodincludes combining an isolated Y641 mutant of EZH2 with a histonesubstrate, a methyl group donor (e.g., SAM), and a test compound,wherein the histone substrate comprises a form of H3-K27 selected fromthe group consisting of monomethylated H3-K27, dimethylated H3-K27, anda combination of monomethylated H3-K27 and dimethylated H3-K27, therebyforming a test mixture; combining an isolated wild-type EZH2 with ahistone substrate, a methyl group donor (e.g., SAM), and a testcompound, wherein the histone substrate comprises a form of H3-K27selected from the group consisting of monomethylated H3-K27,dimethylated H3-K27, and a combination of monomethylated H3-K27 anddimethylated H3-K27, thereby forming a control mixture; performing anassay to detect trimethylation of the histone substrate in each of thetest mixture and the control mixture; calculating the ratio of (a)trimethylation with the Y641 mutant of EZH2 and the test compound (M+)to (b) trimethylation with the Y641 mutant of EZH2 without the testcompound (M−); calculating the ratio of (c) trimethylation withwild-type EZH2 and the test compound (WT+) to (d) trimethylation withwild-type EZH2 without the test compound (WT−); comparing the ratio(a)/(b) with the ratio (c)/(d); and identifying the test compound as aselective inhibitor of the Y641 mutant of EZH2 when the ratio (a)/(b) isless than the ratio (c)/(d). In one embodiment the method furtherincludes taking into account a negative control without test compoundfor either or both of the test mixture and the control mixture.

In some assays, immunological reagents, e.g., antibodies and antigens,are employed. Fluorescence can be utilized in the measurement ofenzymatic activity in some assays. As used herein, “fluorescence” refersto a process through which a molecule emits a photon as a result ofabsorbing an incoming photon of higher energy by the same molecule.Specific methods for assessing the biological activity of the disclosedcompounds are described in the examples.

Administering a compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, polymorph or solvate thereof, to acell or a subject in need thereof results in modulation (i.e.,stimulation or inhibition) of an activity of an intracellular target(e.g., substrate). Several intracellular targets can be modulated withthe compounds of the present invention, including, but not limited to,protein methyltrasferase.

Activating refers to placing a composition of matter (e.g., protein ornucleic acid) in a state suitable for carrying out a desired biologicalfunction. A composition of matter capable of being activated also has anunactivated state. An activated composition of matter may have aninhibitory or stimulatory biological function, or both.

Elevation refers to an increase in a desired biological activity of acomposition of matter (e.g., a protein or a nucleic acid). Elevation mayoccur through an increase in concentration of a composition of matter.

As used herein, “a cell cycle checkpoint pathway” refers to abiochemical pathway that is involved in modulation of a cell cyclecheckpoint. A cell cycle checkpoint pathway may have stimulatory orinhibitory effects, or both, on one or more functions comprising a cellcycle checkpoint. A cell cycle checkpoint pathway is comprised of atleast two compositions of matter, preferably proteins, both of whichcontribute to modulation of a cell cycle checkpoint. A cell cyclecheckpoint pathway may be activated through an activation of one or moremembers of the cell cycle checkpoint pathway. Preferably, a cell cyclecheckpoint pathway is a biochemical signaling pathway.

As used herein, “cell cycle checkpoint regulator” refers to acomposition of matter that can function, at least in part, in modulationof a cell cycle checkpoint. A cell cycle checkpoint regulator may havestimulatory or inhibitory effects, or both, on one or more functionscomprising a cell cycle checkpoint. A cell cycle checkpoint regulatorcan be a protein or not a protein.

Treating cancer or a cell proliferative disorder can result in celldeath, and preferably, cell death results in a decrease of at least 10%in number of cells in a population. More preferably, cell death means adecrease of at least 20%; more preferably, a decrease of at least 30%;more preferably, a decrease of at least 40%; more preferably, a decreaseof at least 50%; most preferably, a decrease of at least 75%. Number ofcells in a population may be measured by any reproducible means. Anumber of cells in a population can be measured by fluorescenceactivated cell sorting (FACS), immunofluorescence microscopy and lightmicroscopy. Methods of measuring cell death are as shown in Li et al.,Proc Natl Acad Sci USA. 100(5): 2674-8, 2003. In an aspect, cell deathoccurs by apoptosis.

Preferably, an effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, is not significantly cytotoxic to normal cells. Atherapeutically effective amount of a compound is not significantlycytotoxic to normal cells if administration of the compound in atherapeutically effective amount does not induce cell death in greaterthan 10% of normal cells. A therapeutically effective amount of acompound does not significantly affect the viability of normal cells ifadministration of the compound in a therapeutically effective amountdoes not induce cell death in greater than 10% of normal cells. In anaspect, cell death occurs by apoptosis.

Contacting a cell with a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, can induce or activate cell death selectively in cancercells. Administering to a subject in need thereof a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, can induce or activate celldeath selectively in cancer cells. Contacting a cell with a compound ofthe present invention, or a pharmaceutically acceptable salt, prodrug,metabolite, polymorph or solvate thereof, can induce cell deathselectively in one or more cells affected by a cell proliferativedisorder. Preferably, administering to a subject in need thereof acompound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, induces celldeath selectively in one or more cells affected by a cell proliferativedisorder.

The present invention relates to a method of treating or preventingcancer by administering a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, to a subject in need thereof, where administration ofthe compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, polymorph or solvate thereof, results in oneor more of the following: prevention of cancer cell proliferation byaccumulation of cells in one or more phases of the cell cycle (e.g. G1,G1/S, G2/M), or induction of cell senescence, or promotion of tumor celldifferentiation; promotion of cell death in cancer cells viacytotoxicity, necrosis or apoptosis, without a significant amount ofcell death in normal cells, antitumor activity in animals with atherapeutic index of at least 2. As used herein, “therapeutic index” isthe maximum tolerated dose divided by the efficacious dose.

One skilled in the art may refer to general reference texts for detaileddescriptions of known techniques discussed herein or equivalenttechniques. These texts include Ausubel et al., Current Protocols inMolecular Biology, John Wiley and Sons, Inc. (2005); Sambrook et al.,Molecular Cloning, A Laboratory Manual (3^(rd) edition), Cold SpringHarbor Press, Cold Spring Harbor, N.Y. (2000); Coligan et al., CurrentProtocols in Immunology, John Wiley & Sons, N.Y.; Enna et al., CurrentProtocols in Pharmacology, John Wiley & Sons, N.Y.; Fingl et al., ThePharmacological Basis of Therapeutics (1975), Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa., 18th edition (1990). Thesetexts can, of course, also be referred to in making or using an aspectof the invention

As used herein, “combination therapy” or “co-therapy” includes theadministration of a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, and at least a second agent as part of a specifictreatment regimen intended to provide the beneficial effect from theco-action of these therapeutic agents. The beneficial effect of thecombination includes, but is not limited to, pharmacokinetic orpharmacodynamic co-action resulting from the combination of therapeuticagents. Administration of these therapeutic agents in combinationtypically is carried out over a defined time period (usually minutes,hours, days or weeks depending upon the combination selected).“Combination therapy” may be, but generally is not, intended toencompass the administration of two or more of these therapeutic agentsas part of separate monotherapy regimens that incidentally andarbitrarily result in the combinations of the present invention.

“Combination therapy” is intended to embrace administration of thesetherapeutic agents in a sequential manner, wherein each therapeuticagent is administered at a different time, as well as administration ofthese therapeutic agents, or at least two of the therapeutic agents, ina substantially simultaneous manner. Substantially simultaneousadministration can be accomplished, for example, by administering to thesubject a single capsule having a fixed ratio of each therapeutic agentor in multiple, single capsules for each of the therapeutic agents.Sequential or substantially simultaneous administration of eachtherapeutic agent can be effected by any appropriate route including,but not limited to, oral routes, intravenous routes, intramuscularroutes, and direct absorption through mucous membrane tissues. Thetherapeutic agents can be administered by the same route or by differentroutes. For example, a first therapeutic agent of the combinationselected may be administered by intravenous injection while the othertherapeutic agents of the combination may be administered orally.Alternatively, for example, all therapeutic agents may be administeredorally or all therapeutic agents may be administered by intravenousinjection. The sequence in which the therapeutic agents are administeredis not narrowly critical.

“Combination therapy” also embraces the administration of thetherapeutic agents as described above in further combination with otherbiologically active ingredients and non-drug therapies (e.g., surgery orradiation treatment). Where the combination therapy further comprises anon-drug treatment, the non-drug treatment may be conducted at anysuitable time so long as a beneficial effect from the co-action of thecombination of the therapeutic agents and non-drug treatment isachieved. For example, in appropriate cases, the beneficial effect isstill achieved when the non-drug treatment is temporally removed fromthe administration of the therapeutic agents, perhaps by days or evenweeks.

A compound of the present invention, or a pharmaceutically acceptablesalt, prodrug, metabolite, analog or derivative thereof, may beadministered in combination with a second chemotherapeutic agent. Thesecond chemotherapeutic agent (also referred to as an anti-neoplasticagent or anti-proliferative agent) can be an alkylating agent; anantibiotic; an anti-metabolite; a detoxifying agent; an interferon; apolyclonal or monoclonal antibody; an EGFR inhibitor; a HER2 inhibitor;a histone deacetylase inhibitor; a hormone; a mitotic inhibitor; an MTORinhibitor; a multi-kinase inhibitor; a serine/threonine kinaseinhibitor; a tyrosine kinase inhibitors; a VEGF/VEGFR inhibitor; ataxane or taxane derivative, an aromatase inhibitor, an anthracycline, amicrotubule targeting drug, a topoisomerase poison drug, an inhibitor ofa molecular target or enzyme (e.g., a kinase or a proteinmethyltransferase), a cytidine analogue drug or any chemotherapeutic,anti-neoplastic or anti-proliferative agent listed inwww.cancer.org/docroot/cdg/cdg_0. asp.

Exemplary alkylating agents include, but are not limited to,cyclophosphamide (Cytoxan; Neosar); chlorambucil (Leukeran); melphalan(Alkeran); carmustine (BiCNU); busulfan (Busulfex); lomustine (CeeNU);dacarbazine (DTIC-Dome); oxaliplatin (Eloxatin); carmustine (Gliadel);ifosfamide (Ifex); mechlorethamine (Mustargen); busulfan (Myleran);carboplatin (Paraplatin); cisplatin (CDDP; Platinol); temozolomide(Temodar); thiotepa (Thioplex); bendamustine (Treanda); or streptozocin(Zanosar).

Exemplary antibiotics include, but are not limited to, doxorubicin(Adriamycin); doxorubicin liposomal (Doxil); mitoxantrone (Novantrone);bleomycin (Blenoxane); daunorubicin (Cerubidine); daunorubicin liposomal(DaunoXome); dactinomycin (Cosmegen); epirubicin (Ellence); idarubicin(Idamycin); plicamycin (Mithracin); mitomycin (Mutamycin); pentostatin(Nipent); or valrubicin (Valstar).

Exemplary anti-metabolites include, but are not limited to, fluorouracil(Adrucil); capecitabine (Xeloda); hydroxyurea (Hydrea); mercaptopurine(Purinethol); pemetrexed (Alimta); fludarabine (Fludara); nelarabine(Arranon); cladribine (Cladribine Novaplus); clofarabine (Clolar);cytarabine (Cytosar-U); decitabine (Dacogen); cytarabine liposomal(DepoCyt); hydroxyurea (Droxia); pralatrexate (Folotyn); floxuridine(FUDR); gemcitabine (Gemzar); cladribine (Leustatin); fludarabine(Oforta); methotrexate (MTX; Rheumatrex); methotrexate (Trexall);thioguanine (Tabloid); TS-1 or cytarabine (Tarabine PFS).

Exemplary detoxifying agents include, but are not limited to, amifostine(Ethyol) or mesna (Mesnex).

Exemplary interferons include, but are not limited to, interferonalfa-2b (Intron A) or interferon alfa-2a (Roferon-A).

Exemplary polyclonal or monoclonal antibodies include, but are notlimited to, trastuzumab (Herceptin); ofatumumab (Arzerra); bevacizumab(Avastin); rituximab (Rittman); cetuximab (Erbitux); panitumumab(Vectibix); tositumomab/iodine131 tositumomab (Bexxar); alemtuzumab(Campath); ibritumomab (Zevalin; In-111; Y-90 Zevalin); gemtuzumab(Mylotarg); eculizumab (Soliris) ordenosumab.

Exemplary EGFR inhibitors include, but are not limited to, gefitinib(Iressa); lapatinib (Tykerb); cetuximab (Erbitux); erlotinib (Tarceva);panitumumab (Vectibix); PKI-166; canertinib (CI-1033); matuzumab(Emd7200) or EKB-569.

Exemplary HER2 inhibitors include, but are not limited to, trastuzumab(Herceptin); lapatinib (Tykerb) or AC-480.

Histone Deacetylase Inhibitors include, but are not limited to,vorinostat (Zolinza).

Exemplary hormones include, but are not limited to, tamoxifen (Soltamox;Nolvadex); raloxifene (Evista); megestrol (Megace); leuprolide (Lupron;Lupron Depot; Eligard; Viadur); fulvestrant (Faslodex); letrozole(Femara); triptorelin (Trelstar LA; Trelstar Depot); exemestane(Aromasin); goserelin (Zoladex); bicalutamide (Casodex); anastrozole(Arimidex); fluoxymesterone (Androxy; Halotestin); medroxyprogesterone(Provera; Depo-Provera); estramustine (Emcyt); flutamide (Eulexin);toremifene (Fareston); degarelix (Firmagon); nilutamide (Nilandron);abarelix (Plenaxis); or testolactone (Teslac).

Exemplary mitotic inhibitors include, but are not limited to, paclitaxel(Taxol; Onxol; Abraxane); docetaxel (Taxotere); vincristine (Oncovin;Vincasar PFS); vinblastine (Velban); etoposide (Toposar; Etopophos;VePesid); teniposide (Vumon); ixabepilone (Ixempra); nocodazole;epothilone; vinorelbine (Navelbine); camptothecin (CPT); irinotecan(Camptosar); topotecan (Hycamtin); amsacrine or lamellarin D (LAM-D).

Exemplary MTOR inhibitors include, but are not limited to, everolimus(Afinitor) or temsirolimus (Torisel); rapamune, ridaforolimus; orAP23573.

Exemplary multi-kinase inhibitors include, but are not limited to,sorafenib (Nexavar); sunitinib (Sutent); BIBW 2992; E7080; Zd6474;PKC-412; motesanib; or AP24534.

Exemplary serine/threonine kinase inhibitors include, but are notlimited to, ruboxistaurin; eril/easudil hydrochloride; flavopiridol;seliciclib (CYC202; Roscovitrine); SNS-032 (BMS-387032); Pkc412;bryostatin; KAI-9803; SF1126; VX-680; Azd1152; Arry-142886 (AZD-6244);SCIO-469; GW681323; CC-401; CEP-1347 or PD 332991.

Exemplary tyrosine kinase inhibitors include, but are not limited to,erlotinib (Tarceva); gefitinib (Iressa); imatinib (Gleevec); sorafenib(Nexavar); sunitinib (Sutent); trastuzumab (Herceptin); bevacizumab(Avastin); rituximab (Rituxan); lapatinib (Tykerb); cetuximab (Erbitux);panitumumab (Vectibix); everolimus (Afinitor); alemtuzumab (Campath);gemtuzumab (Mylotarg); temsirolimus (Torisel); pazopanib (Votrient);dasatinib (Sprycel); nilotinib (Tasigna); vatalanib (Ptk787; ZK222584);CEP-701; SU5614; MLN518; XL999; VX-322; Azd0530; BMS-354825; SKI-606CP-690; AG-490; WHI-P154; WHI-P131; AC-220; or AMG888.

Exemplary VEGF/VEGFR inhibitors include, but are not limited to,bevacizumab (Avastin); sorafenib (Nexavar); sunitinib (Sutent);ranibizumab; pegaptanib; or vandetinib.

Exemplary microtubule targeting drugs include, but are not limited to,paclitaxel, docetaxel, vincristin, vinblastin, nocodazole, epothilonesand navelbine.

Exemplary topoisomerase poison drugs include, but are not limited to,teniposide, etoposide, adriamycin, camptothecin, daunorubicin,dactinomycin, mitoxantrone, amsacrine, epirubicin and idarubicin.

Exemplary taxanes or taxane derivatives include, but are not limited to,paclitaxel and docetaxol.

Exemplary general chemotherapeutic, anti-neoplastic, anti-proliferativeagents include, but are not limited to, altretamine (Hexalen);isotretinoin (Accutane; Amnesteem; Claravis; Sotret); tretinoin(Vesanoid); azacitidine (Vidaza); bortezomib (Velcade) asparaginase(Elspar); levamisole (Ergamisol); mitotane (Lysodren); procarbazine(Matulane); pegaspargase (Oncaspar); denileukin diftitox (Ontak);porfimer (Photofrin); aldesleukin (Proleukin); lenalidomide (Revlimid);bexarotene (Targretin); thalidomide (Thalomid); temsirolimus (Torisel);arsenic trioxide (Trisenox); verteporfin (Visudyne); mimosine(Leucenol); (1M tegafur—0.4 M 5-chloro-2,4-dihydroxypyrimidine—1 Mpotassium oxonate) or lovastatin.

In another aspect, the second chemotherapeutic agent can be a cytokinesuch as G-CSF (granulocyte colony stimulating factor). In anotheraspect, a compound of the present invention, or a pharmaceuticallyacceptable salt, prodrug, metabolite, analog or derivative thereof, maybe administered in combination with radiation therapy. Radiation therapycan also be administered in combination with a compound of the presentinvention and another chemotherapeutic agent described herein as part ofa multiple agent therapy. In yet another aspect, a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug,metabolite, analog or derivative thereof, may be administered incombination with standard chemotherapy combinations such as, but notrestricted to, CMF (cyclophosphamide, methotrexate and 5-fluorouracil),CAF (cyclophosphamide, adriamycin and 5-fluorouracil), AC (adriamycinand cyclophosphamide), FEC (5-fluorouracil, epirubicin, andcyclophosphamide), ACT or ATC (adriamycin, cyclophosphamide, andpaclitaxel), rituximab, Xeloda (capecitabine), Cisplatin (CDDP),Carboplatin, TS-1 (tegafur, gimestat and otastat potassium at a molarratio of 1:0.4:1), Camptothecin-11 (CPT-11, Irinotecan or Camptosar™) orCMFP (cyclophosphamide, methotrexate, 5-fluorouracil and prednisone).

In preferred embodiments, a compound of the present invention, or apharmaceutically acceptable salt, prodrug, metabolite, polymorph orsolvate thereof, may be administered with an inhibitor of an enzyme,such as a receptor or non-receptor kinase. Receptor and non-receptorkinases are, for example, tyrosine kinases or serine/threonine kinases.Kinase inhibitors described herein are small molecules, polynucleicacids, polypeptides, or antibodies.

Exemplary kinase inhibitors include, but are not limited to, Bevacizumab(targets VEGF), BIBW 2992 (targets EGFR and Erb2), Cetuximab/Erbitux(targets Erb1), Imatinib/Gleevic (targets Bcr-Abl), Trastuzumab (targetsErb2), Gefitinib/Iressa (targets EGFR), Ranibizumab (targets VEGF),Pegaptanib (targets VEGF), Erlotinib/Tarceva (targets Erb1), Nilotinib(targets Bcr-Abl), Lapatinib (targets Erb1 and Erb2/Her2),GW-572016/lapatinib ditosylate (targets HER2/Erb2), Panitumumab/Vectibix(targets EGFR), Vandetinib (targets RET/VEGFR), E7080 (multiple targetsincluding RET and VEGFR), Herceptin (targets HER2/Erb2), PM-166 (targetsEGFR), Canertinib/CI-1033 (targets EGFR), Sunitinib/SU-11464/Sutent(targets EGFR and FLT3), Matuzumab/Emd7200 (targets EGFR), EKB-569(targets EGFR), Zd6474 (targets EGFR and VEGFR), PKC-412 (targets VEGRand FLT3), Vatalanib/Ptk787/ZK222584 (targets VEGR), CEP-701 (targetsFLT3), SU5614 (targets FLT3), MLN518 (targets FLT3), XL999 (targetsFLT3), VX-322 (targets FLT3), Azd0530 (targets SRC), BMS-354825 (targetsSRC), SM-606 (targets SRC), CP-690 (targets JAK), AG-490 (targets JAK),WHI-P154 (targets JAK), WHI-P131 (targets JAK), sorafenib/Nexavar(targets RAF kinase, VEGFR-1, VEGFR-2, VEGFR-3, PDGFR-β, MT, FLT-3, andRET), Dasatinib/Sprycel (BCR/ABL and Src), AC-220 (targets Flt3), AC-480(targets all HER proteins, “panHER”), Motesanib diphosphate (targetsVEGF1-3, PDGFR, and c-kit), Denosumab (targets RANKL, inhibits SRC),AMG888 (targets HER3), and AP24534 (multiple targets including Flt3).

Exemplary serine/threonine kinase inhibitors include, but are notlimited to, Rapamune (targets mTOR/FRAP1), Deforolimus (targets mTOR),Certican/Everolimus (targets mTOR/FRAP1), AP23573 (targets mTOR/FRAP1),Eril/Fasudil hydrochloride (targets RHO), Flavopiridol (targets CDK),Seliciclib/CYC₂₀₂/Roscovitrine (targets CDK), SNS-032/BMS-387032(targets CDK), Ruboxistaurin (targets PKC), Pkc412 (targets PKC),Bryostatin (targets PKC), KAI-9803 (targets PKC), SF1126 (targets PI3K),VX-680 (targets Aurora kinase), Azd1152 (targets Aurora kinase),Any-142886/AZD-6244 (targets MAP/MEK), SCID-469 (targets MAP/MEK),GW681323 (targets MAP/MEK), CC-401 (targets JNK), CEP-1347 (targetsJNK), and PD 332991 (targets CDK).

The disorder in which EZH2-mediated protein methylation plays a part canbe a neurological disease. The compound of this invention can thus alsobe used for treating neurologic diseases such as epilepsy,schizophrenia, bipolar disorder or other psychological and/orpsychiatric disorders, neuropathies, skeletal muscle atrophy, andneurodegenerative diseases, e.g., a neurodegenerative disease. Exemplaryneurodegenerative diseases include: Alzheimer's, Amyotrophic LateralSclerosis (ALS), and Parkinson's disease. Another class ofneurodegenerative diseases includes diseases caused at least in part byaggregation of poly-glutamine. Diseases of this class include:Huntington's Diseases, Spinalbulbar Muscular Atrophy (SBMA or Kennedy'sDisease) Dentatorubropallidoluysian Atrophy (DRPLA), SpinocerebellarAtaxia 1 (SCA1), Spinocerebellar Ataxia 2 (SCA2), Machado-Joseph Disease(MJD; SCA3), Spinocerebellar Ataxia 6 (SCA6), Spinocerebellar Ataxia 7(SCAT), and Spinocerebellar Ataxia 12 (SCA12).

Any other disease in which epigenetic methylation, which is mediated byEZH2, plays a role may be treatable or preventable using compounds andmethods described herein

4. Pharmaceutical Compositions

The present invention also provides pharmaceutical compositionscomprising a compound of Formulae (I), (Ia), and (Ib) in combinationwith at least one pharmaceutically acceptable excipient or carrier.

A “pharmaceutical composition” is a formulation containing the compoundsof the present invention in a form suitable for administration to asubject. In one embodiment, the pharmaceutical composition is in bulk orin unit dosage form. The unit dosage form is any of a variety of forms,including, for example, a capsule, an IV bag, a tablet, a single pump onan aerosol inhaler or a vial. The quantity of active ingredient (e.g., aformulation of the disclosed compound or salt, hydrate, solvate orisomer thereof) in a unit dose of composition is an effective amount andis varied according to the particular treatment involved. One skilled inthe art will appreciate that it is sometimes necessary to make routinevariations to the dosage depending on the age and condition of thepatient. The dosage will also depend on the route of administration. Avariety of routes are contemplated, including oral, pulmonary, rectal,parenteral, transdermal, subcutaneous, intravenous, intramuscular,intraperitoneal, inhalational, buccal, sublingual, intrapleural,intrathecal, intranasal, and the like. Dosage forms for the topical ortransdermal administration of a compound of this invention includepowders, sprays, ointments, pastes, creams, lotions, gels, solutions,patches and inhalants. In one embodiment, the active compound is mixedunder sterile conditions with a pharmaceutically acceptable carrier, andwith any preservatives, buffers, or propellants that are required.

As used herein, the phrase “pharmaceutically acceptable” refers to thosecompounds, anions, cations, materials, compositions, carriers, and/ordosage forms which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of human beings and animalswithout excessive toxicity, irritation, allergic response, or otherproblem or complication, commensurate with a reasonable benefit/riskratio.

“Pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic and neither biologically nor otherwise undesirable, andincludes excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

A pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. Examples of routesof administration include parenteral, e.g., intravenous, intradermal,subcutaneous, oral (e.g., inhalation), transdermal (topical), andtransmucosal administration. Solutions or suspensions used forparenteral, intradermal, or subcutaneous application can include thefollowing components: a sterile diluent such as water for injection,saline solution, fixed oils, polyethylene glycols, glycerine, propyleneglycol or other synthetic solvents; antibacterial agents such as benzylalcohol or methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfate; chelating agents such as ethylenediaminetetraacetic acid;buffers such as acetates, citrates or phosphates, and agents for theadjustment of tonicity such as sodium chloride or dextrose. The pH canbe adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide. The parenteral preparation can be enclosed in ampoules,disposable syringes or multiple dose vials made of glass or plastic.

A compound or pharmaceutical composition of the invention can beadministered to a subject in many of the well-known methods currentlyused for chemotherapeutic treatment. For example, for treatment ofcancers, a compound of the invention may be injected directly intotumors, injected into the blood stream or body cavities or taken orallyor applied through the skin with patches. The dose chosen should besufficient to constitute effective treatment but not so high as to causeunacceptable side effects. The state of the disease condition (e.g.,cancer, precancer, and the like) and the health of the patient shouldpreferably be closely monitored during and for a reasonable period aftertreatment.

The term “therapeutically effective amount”, as used herein, refers toan amount of a pharmaceutical agent to treat, ameliorate, or prevent anidentified disease or condition, or to exhibit a detectable therapeuticor inhibitory effect. The effect can be detected by any assay methodknown in the art. The precise effective amount for a subject will dependupon the subject's body weight, size, and health; the nature and extentof the condition; and the therapeutic or combination of therapeuticsselected for administration. Therapeutically effective amounts for agiven situation can be determined by routine experimentation that iswithin the skill and judgment of the clinician. In a preferred aspect,the disease or condition to be treated is cancer. In another aspect, thedisease or condition to be treated is a cell proliferative disorder.

For any compound, the therapeutically effective amount can be estimatedinitially either in cell culture assays, e.g., of neoplastic cells, orin animal models, usually rats, mice, rabbits, dogs, or pigs. The animalmodel may also be used to determine the appropriate concentration rangeand route of administration. Such information can then be used todetermine useful doses and routes for administration in humans.Therapeutic/prophylactic efficacy and toxicity may be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., ED₅₀ (the dose therapeutically effective in 50% of thepopulation) and LD₅₀ (the dose lethal to 50% of the population). Thedose ratio between toxic and therapeutic effects is the therapeuticindex, and it can be expressed as the ratio, LD₅₀/ED₅₀. Pharmaceuticalcompositions that exhibit large therapeutic indices are preferred. Thedosage may vary within this range depending upon the dosage formemployed, sensitivity of the patient, and the route of administration.

Dosage and administration are adjusted to provide sufficient levels ofthe active agent(s) or to maintain the desired effect. Factors which maybe taken into account include the severity of the disease state, generalhealth of the subject, age, weight, and gender of the subject, diet,time and frequency of administration, drug combination(s), reactionsensitivities, and tolerance/response to therapy. Long-actingpharmaceutical compositions may be administered every 3 to 4 days, everyweek, or once every two weeks depending on half-life and clearance rateof the particular formulation.

The pharmaceutical compositions containing active compounds of thepresent invention may be manufactured in a manner that is generallyknown, e.g., by means of conventional mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping, orlyophilizing processes. Pharmaceutical compositions may be formulated ina conventional manner using one or more pharmaceutically acceptablecarriers comprising excipients and/or auxiliaries that facilitateprocessing of the active compounds into preparations that can be usedpharmaceutically. Of course, the appropriate formulation is dependentupon the route of administration chosen.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. For intravenous administration, suitablecarriers include physiological saline, bacteriostatic water, CremophorEL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In allcases, the composition must be sterile and should be fluid to the extentthat easy syringeability exists. It must be stable under the conditionsof manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (for example, glycerol, propylene glycol, andliquid polyethylene glycol, and the like), and suitable mixturesthereof. The proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prevention of the action of microorganisms can be achieved by variousantibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, it will be preferable to include isotonic agents, for example,sugars, polyalcohols such as manitol and sorbitol, and sodium chloridein the composition. Prolonged absorption of the injectable compositionscan be brought about by including in the composition an agent whichdelays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the activecompound in the required amount in an appropriate solvent with one or acombination of ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the active compound into a sterile vehicle that contains abasic dispersion medium and the required other ingredients from thoseenumerated above. In the case of sterile powders for the preparation ofsterile injectable solutions, methods of preparation are vacuum dryingand freeze-drying that yields a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof

Oral compositions generally include an inert diluent or an ediblepharmaceutically acceptable carrier. They can be enclosed in gelatincapsules or compressed into tablets. For the purpose of oral therapeuticadministration, the active compound can be incorporated with excipientsand used in the form of tablets, troches, or capsules. Oral compositionscan also be prepared using a fluid carrier for use as a mouthwash,wherein the compound in the fluid carrier is applied orally and swishedand expectorated or swallowed. Pharmaceutically compatible bindingagents, and/or adjuvant materials can be included as part of thecomposition. The tablets, pills, capsules, troches and the like cancontain any of the following ingredients, or compounds of a similarnature: a binder such as microcrystalline cellulose, gum tragacanth orgelatin; an excipient such as starch or lactose, a disintegrating agentsuch as alginic acid, Primogel, or corn starch; a lubricant such asmagnesium stearate or Sterotes; a glidant such as colloidal silicondioxide; a sweetening agent such as sucrose or saccharin; or a flavoringagent such as peppermint, methyl salicylate, or orange flavoring.

For administration by inhalation, the compounds are delivered in theform of an aerosol spray from pressured container or dispenser, whichcontains a suitable propellant, e.g., a gas such as carbon dioxide, or anebulizer.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art, and include, forexample, for transmucosal administration, detergents, bile salts, andfusidic acid derivatives. Transmucosal administration can beaccomplished through the use of nasal sprays or suppositories. Fortransdermal administration, the active compounds are formulated intoointments, salves, gels, or creams as generally known in the art.

The active compounds can be prepared with pharmaceutically acceptablecarriers that will protect the compound against rapid elimination fromthe body, such as a controlled release formulation, including implantsand microencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

It is especially advantageous to formulate oral or parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subject tobe treated; each unit containing a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on the unique characteristics of the active compound and theparticular therapeutic effect to be achieved.

In therapeutic applications, the dosages of the pharmaceuticalcompositions used in accordance with the invention vary depending on theagent, the age, weight, and clinical condition of the recipient patient,and the experience and judgment of the clinician or practitioneradministering the therapy, among other factors affecting the selecteddosage. Generally, the dose should be sufficient to result in slowing,and preferably regressing, the growth of the tumors and also preferablycausing complete regression of the cancer. Dosages can range from about0.01 mg/kg per day to about 5000 mg/kg per day. In preferred aspects,dosages can range from about 1 mg/kg per day to about 1000 mg/kg perday. In an aspect, the dose will be in the range of about 0.1 mg/day toabout 50 g/day; about 0.1 mg/day to about 25 g/day; about 0.1 mg/day toabout 10 g/day; about 0.1 mg to about 3 g/day; or about 0.1 mg to about1 g/day, in single, divided, or continuous doses (which dose may beadjusted for the patient's weight in kg, body surface area in m², andage in years). An effective amount of a pharmaceutical agent is thatwhich provides an objectively identifiable improvement as noted by theclinician or other qualified observer. For example, regression of atumor in a patient may be measured with reference to the diameter of atumor. Decrease in the diameter of a tumor indicates regression.Regression is also indicated by failure of tumors to reoccur aftertreatment has stopped. As used herein, the term “dosage effectivemanner” refers to amount of an active compound to produce the desiredbiological effect in a subject or cell.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

The compounds of the present invention are capable of further formingsalts. All of these forms are also contemplated within the scope of theclaimed invention.

As used herein, “pharmaceutically acceptable salts” refer to derivativesof the compounds of the present invention wherein the parent compound ismodified by making acid or base salts thereof. Examples ofpharmaceutically acceptable salts include, but are not limited to,mineral or organic acid salts of basic residues such as amines, alkalior organic salts of acidic residues such as carboxylic acids, and thelike. The pharmaceutically acceptable salts include the conventionalnon-toxic salts or the quaternary ammonium salts of the parent compoundformed, for example, from non-toxic inorganic or organic acids. Forexample, such conventional non-toxic salts include, but are not limitedto, those derived from inorganic and organic acids selected from2-acetoxybenzoic, 2-hydroxyethane sulfonic, acetic, ascorbic, benzenesulfonic, benzoic, bicarbonic, carbonic, citric, edetic, ethanedisulfonic, 1,2-ethane sulfonic, fumaric, glucoheptonic, gluconic,glutamic, glycolic, glycollyarsanilic, hexylresorcinic, hydrabamic,hydrobromic, hydrochloric, hydroiodic, hydroxymaleic, hydroxynaphthoic,isethionic, lactic, lactobionic, lauryl sulfonic, maleic, malic,mandelic, methane sulfonic, napsylic, nitric, oxalic, pamoic,pantothenic, phenylacetic, phosphoric, polygalacturonic, propionic,salicyclic, stearic, subacetic, succinic, sulfamic, sulfanilic,sulfuric, tannic, tartaric, toluene sulfonic, and the commonly occurringamine acids, e.g., glycine, alanine, phenylalanine, arginine, etc.

Other examples of pharmaceutically acceptable salts include hexanoicacid, cyclopentane propionic acid, pyruvic acid, malonic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonicacid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid,camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylicacid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylaceticacid, muconic acid, and the like. The present invention also encompassessalts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

It should be understood that all references to pharmaceuticallyacceptable salts include solvent addition forms (solvates) or crystalforms (polymorphs) as defined herein, of the same salt.

The compounds of the present invention can also be prepared as esters,for example, pharmaceutically acceptable esters. For example, acarboxylic acid function group in a compound can be converted to itscorresponding ester, e.g., a methyl, ethyl or other ester. Also, analcohol group in a compound can be converted to its corresponding ester,e.g., acetate, propionate or other ester.

The compounds of the present invention can also be prepared as prodrugs,for example, pharmaceutically acceptable prodrugs. The terms “pro-drug”and “prodrug” are used interchangeably herein and refer to any compoundwhich releases an active parent drug in vivo. Since prodrugs are knownto enhance numerous desirable qualities of pharmaceuticals (e.g.,solubility, bioavailability, manufacturing, etc.), the compounds of thepresent invention can be delivered in prodrug form. Thus, the presentinvention is intended to cover prodrugs of the presently claimedcompounds, methods of delivering the same and compositions containingthe same. “Prodrugs” are intended to include any covalently bondedcarriers that release an active parent drug of the present invention invivo when such prodrug is administered to a subject. Prodrugs in thepresent invention are prepared by modifying functional groups present inthe compound in such a way that the modifications are cleaved, either inroutine manipulation or in vivo, to the parent compound. Prodrugsinclude compounds of the present invention wherein a hydroxy, amino,sulfhydryl, carboxy or carbonyl group is bonded to any group that may becleaved in vivo to form a free hydroxyl, free amino, free sulfhydryl,free carboxy or free carbonyl group, respectively.

Examples of prodrugs include, but are not limited to, esters (e.g.,acetate, dialkylaminoacetates, formates, phosphates, sulfates andbenzoate derivatives) and carbamates (e.g., N,N-dimethylaminocarbonyl)of hydroxy functional groups, esters (e.g., ethyl esters,morpholinoethanol esters) of carboxyl functional groups, N-acylderivatives (e.g., N-acetyl) N-Mannich bases, Schiff bases andenaminones of amino functional groups, oximes, acetals, ketals and enolesters of ketone and aldehyde functional groups in compounds of theinvention, and the like, See Bundegaard, H., Design of Prodrugs, p 1-92,Elesevier, New York-Oxford (1985).

The compounds, or pharmaceutically acceptable salts, esters or prodrugsthereof, are administered orally, nasally, transdermally, pulmonary,inhalationally, buccally, sublingually, intraperintoneally,subcutaneously, intramuscularly, intravenously, rectally,intrapleurally, intrathecally and parenterally. In one embodiment, thecompound is administered orally. One skilled in the art will recognizethe advantages of certain routes of administration.

The dosage regimen utilizing the compounds is selected in accordancewith a variety of factors including type, species, age, weight, sex andmedical condition of the patient; the severity of the condition to betreated; the route of administration; the renal and hepatic function ofthe patient; and the particular compound or salt thereof employed. Anordinarily skilled physician or veterinarian can readily determine andprescribe the effective amount of the drug required to prevent, counter,or arrest the progress of the condition.

Techniques for formulation and administration of the disclosed compoundsof the invention can be found in Remington: the Science and Practice ofPharmacy, 19th edition, Mack Publishing Co., Easton, Pa. (1995). In anembodiment, the compounds described herein, and the pharmaceuticallyacceptable salts thereof, are used in pharmaceutical preparations incombination with a pharmaceutically acceptable carrier or diluent.Suitable pharmaceutically acceptable carriers include inert solidfillers or diluents and sterile aqueous or organic solutions. Thecompounds will be present in such pharmaceutical compositions in amountssufficient to provide the desired dosage amount in the range describedherein.

All percentages and ratios used herein, unless otherwise indicated, areby weight. Other features and advantages of the present invention areapparent from the different examples. The provided examples illustratedifferent components and methodology useful in practicing the presentinvention. The examples do not limit the claimed invention. Based on thepresent disclosure the skilled artisan can identify and employ othercomponents and methodology useful for practicing the present invention.

In the synthetic schemes described herein, compounds may be drawn withone particular configuration for simplicity. Such particularconfigurations are not to be construed as limiting the invention to oneor another isomer, tautomer, regioisomer or stereoisomer, nor does itexclude mixtures of isomers, tautomers, regioisomers or stereoisomers.

Compounds designed, selected and/or optimized by methods describedabove, once produced, can be characterized using a variety of assaysknown to those skilled in the art to determine whether the compoundshave biological activity. For example, the molecules can becharacterized by conventional assays, including but not limited to thoseassays described below, to determine whether they have a predictedactivity, binding activity and/or binding specificity.

Furthermore, high-throughput screening can be used to speed up analysisusing such assays. As a result, it can be possible to rapidly screen themolecules described herein for activity, using techniques known in theart. General methodologies for performing high-throughput screening aredescribed, for example, in Devlin (1998) High Throughput Screening,Marcel Dekker; and U.S. Pat. No. 5,763,263. High-throughput assays canuse one or more different assay techniques including, but not limitedto, those described below.

All publications and patent documents cited herein are incorporatedherein by reference as if each such publication or document wasspecifically and individually indicated to be incorporated herein byreference. Citation of publications and patent documents is not intendedas an admission that any is pertinent prior art, nor does it constituteany admission as to the contents or date of the same. The inventionhaving now been described by way of written description, those of skillin the art will recognize that the invention can be practiced in avariety of embodiments and that the foregoing description and examplesbelow are for purposes of illustration and not limitation of the claimsthat follow.

5. Examples

General Procedure for Suzuki Coupling:

To a stirred solution of bromo compound (1 equiv.) and boronate ester(1.2 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.)was added and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. Reaction masswas heated at 100° C. for 2 h. On completion, reaction mixture wasdiluted with water and extracted with 10% MeOH/DCM. Combined organiclayers were dried over Na₂SO₄ and solvent removed under reduced pressureto afford crude material which was purified by column chromatographyover silica gel to afford desired compound (yield 75-90%).

General Procedure for Boc Deprotection:

A stirred solution of Boc protected amine (1 mmol) in DCM (5 mL) wascooled to 0° C. and TFA (2 mL) was added to it. Reaction mass wasstirred at rt for 1 h. On completion, reaction was concentrated todryness. Residue was basified with aqueous sodium bicarbonate till pH 8and aqueous layer extracted with 20% MeOH/DCM. Combined organic layerswere dried over sodium sulfate and concentrated to afford desiredcompound.

General Procedure for PYBOP Coupling:

To a solution of acid (1 equiv.) and amine (2 equiv.) in DMSO (3 mL for1 mmol), PyBOP (1.5 equiv.) was added and reaction stirred at roomtemperature for overnight. On completion, water was added and solid thatprecipitates out was filtered and washed with water. Then this solid wasstirred with acetonitrile for 10 min and filtered again to obtain puredesired compound.

General Procedure for Alkylation ((Reductive Amination with Aldehydes):

To a stirred solution of starting material (1 equiv.) and appropriatealdehyde (1.5 equiv.) in appropriate solvent (5 mL for 0.3 mmol; MeOH orDCE), acetic acid (1 equiv.) was added and reaction stirred at rt. Thenreducing agent (1 equiv.; NaBH₃CN or Na(OAc)₃BH) was added and reactionstirred overnight. On completion, solvent was removed under reducedpressure and residue purified by column chromatography over silica gelor as specified affording desired compound.

General Procedure for Methylation (Reductive Amination with FormalinSolution):

To a stirred solution of secondary amine (1 equiv.) in methanol (5 mLfor 1 mmol), formalin solution (37-40% sol, 10 equiv.) was added at 5°C. and reaction stirred for 10 min. Then NaBH₃CN (3 equiv.) was addedand reaction stirred at rt for 1 h. On completion, methanol was removedunder reduced pressure; water was added to the residue and aqueous phaseextracted with 10% MeOH/DCM. Combined organic layers were dried oversodium sulfate and concentrated to obtain crude material which waspurified by column chromatography or prep HPLC.

General Procedure Reaction of Benzyl Bromides

To a stirred solution of respective amino-alcohol (3 equiv.) in dry THF,NaH (3 equiv) was added at 0° C. Resulting reaction mass was stirred for30 minutes. To this a solution of the benzyl bromide (1 equiv.) in THFwas added at 0° C. and reaction mass stirred at room temperature for 1h. On completion, reaction was quenched with water and extracted with10% MeOH/DCM. Combined organic layer was dried over sodium sulfate andconcentrated to give crude material which was purified by preparativeHPLC to afford final target molecules as TFA salts.

Procedure for Pd—C Reduction:

To a stirred solution of starting material (0.1 g) in MeOH (5 mL) wasadded 10% Pd/C (0.1 g) and reaction stirred at room temperature underhydrogen (balloon pressure) for 4 h. On completion, the reaction mixturewas filtered through a celite bed and the filtrate concentrated underreduce pressure to a obtain crude solid which was purified by solventwashing to afford final target molecules as TFA salts.

Example 1

The compounds listed in Tables 1-6 were synthesized by reaction schemesdepicted in the general schemes above or by methods described below.

Synthesis of Compound A-1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one

3-Cyano-2,4-dimethyl-2-hydroxypyridine (0.3 g, 2.0 mmol) was dissolvedin MeOH (5 mL), to which catalytic amount of Raney Ni and of aqueous NH₃(0.3 mL) were added and the reaction mixture was stirred under hydrogenpressure (bladder pressure) for 3-4 h. After completion of the reaction,catalyst was filtered off and the filtrate was concentrated underreduced pressure. The residue was thoroughly dried to provide thedesired product (0.3 g, quantitative yield).

Step 2: Synthesis of ethyl6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A stirred solution of 1H-pyrazol-3-amine (45 g, 542 mmol) in acetic acid(297 mL) and water (900 mL) was cooled to 0° C. and diethyl oxaloacetatesodium salt (113.85 g, 542.16 mmol) was added to it. Resulting solutionwas heated at 100° C. for overnight. After completion of reaction, solidwas filtered and dried to obtain the desired intermediate (25 g, 22%).

Step 3: Synthesis of ethyl6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (25 g, 120mmol) was suspended in acetonitrile (250 mL) and POBr₃ (69.56 g, 241.54mmol) was added to it. The reaction mixture was refluxed for 6 h. Oncompletion of reaction, acetonitrile was removed under reduced pressureand residue neutralized with saturated NaHCO₃ solution was added to it.Extraction was carried out using ethyl acetate; the combined organiclayers were washed with water, brine and dried over anhydrous Na₂SO₄;filtered and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography to provide the desiredintermediate (25 g, 77%).

Step 4: Synthesis of ethyl6-bromo-1-cyclopentyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (3 g, 11.15 mmol)was dissolved in acetonitrile (30 mL) and K₂CO₃ (1.85 g, 13.38 mmol) andcyclopentyl bromide (3.35 g, 22.30 mmol) was added to it. The reactionmixture was refluxed for 3 h. On completion, acetonitrile was removedunder reduced pressure and water was added to it. Extraction was carriedout using ethyl acetate; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄. Solvent was removed underreduced pressure and residue was purified by silica gel columnchromatography to obtain the desired intermediate (1.3 g, 50%).

Step 5: Synthesis of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (0.308 g, 7.72 mmol) was added to a solution of ethyl6-bromo-1-cyclopentyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1.3 g,3.8 mmol) in EtOH (10 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand it was acidified using 10% citric acid solution. Extraction wascarried out using ethyl acetate; the combined organic layers were washedwith water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure. The crude acid (1 g, 3.23 mmol) wasthen dissolved in DMSO (10 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.98 g, 6.47 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (2.52 g, 4.85 mmol) was added to it and stirring wascontinued overnight. After completion of the reaction, reaction mass waspoured into ice to obtain solid, which was filtered and washed withacetonitrile followed by ether to provide the desired intermediate (1 g,70%).

Step 6: Synthesis of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.25 g, 0.56 mmol), boronic ester (0.163 g, 0.620 mmol) and Pd(PPh₃)₄(0.065 g, 0.056 mmol) in 1,4-dioxane (3 mL) was purged with argon for 10min. Then, 2 M Na₂CO₃ solution (0.216 g, 2.03 mmol) in water was addedto it and again argon was purged through it for 10 min. The reactionmixture was stirred at 100° C. for 1 h. After completion of thereaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound (72% yield). LCMS: 541.30 (M+1)⁺; HPLC: 99.81%(@ 254 nm) (R_(t); 5.464); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H),8.95 (t, 1H, J=5.2 Hz), 8.36 (s, 1H), 8.20 (d, 2H, J=8.4 Hz), 8.14 (s,1H), 7.48 (d, 2H, J=7.6 Hz), 5.89 (s, 1H), 5.51-5.48 (m, 1H), 4.40 (d,2H, J=4.8 Hz), 3.60-3.58 (m, 4H), 3.54 (s, 2H), 2.38 (bs, 4H), 2.21 (s,3H), 2.18-2.15 (s, 2H), 2.12 (s, 3H), 2.08-2.05 (m, 2H), 1.94-1.92 (m,2H), 1.76-1.73 (m, 2H).

Synthesis of Compound A-2:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl1-isopropyl-6-(trifluoromethylsulfonyloxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Trifluoromethanesulfonic anhydride (5.23 g, 18.5 mmol) was added to acooled solution of ethyl6-hydroxy-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (4.2 g,16.9 mmol) and pyridine (1.46 g, 18.5 mmol) in DCM (60 mL); and it wasallowed to stirred at room temperature for 2 h. After completion of thereaction, saturated NaHCO₃ solution (60 mL) was added to it andextraction was carried out using DCM (30 mL×3). The combined organiclayers were washed with water (100 mL); brine (100 mL); dried overanhydrous Na₂SO₄; filtered and concentrated under reduced pressure. Theresidue was finally purified by silica gel column chromatography toprovide pure the desired intermediate (5 g, 77.8% yield).

Step 2: Synthesis of ethyl6-(4-(hydroxymethyl)phenyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A solution of ethyl1-isopropyl-6-(trifluoromethylsulfonyloxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(1 g, 2.6 mmol), 4-(hydroxymethyl) phenyl boronic acid (0.438 g, 2.88mmol) and Pd(PPh₃)₄ (0.303 g, 0.262 mmol) in 1,4-dioxane (15 mL) waspurged with argon for 10 min. Then, 2M Na₂CO₃ solution (4.5 mL, 9.44mmol) was added to it and again argon was purged through it for 10 min.The reaction mixture was stirred at 100° C. for 1 h. After completion ofthe reaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound.

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a solution of ethyl6-(4-(hydroxymethyl)phenyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.81 g, 2.38 mmol) in EtOH (15 mL), aqueous NaOH (0.47 g, 11.94 mmol)was added and reaction mixture stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand it was acidified using 10% citric acid solution. Extraction wascarried out using 5% MeOH/DCM. The combined organic layers were washedwith water & dried over anhydrous Na₂SO₄, filtered and concentratedunder reduced pressure to afford compound which was pure enough to useas such for next step. A solution of acid compound (0.61 g, 1.96 mmol)and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.596 g, 3.92 mmol)in DMSO (5 mL) and was stirred at room temperature for 15 min. ThenPYBOP (1.52 g, 2.94 mmol) was added to it and stirring was continued for12 hr. After completion of the reaction, saturated NaHCO₃ solution wasadded to it and extraction was carried out using 20% MeOH/DCM. Thecombined organic layers were washed with water, dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to afford thedesired compound as white solid (0.71 g, 81.4%).

Step 4: Synthesis of6-(4-(bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.45 g, 1.01 mmol) in a DCM (10 mL) was added PPh₃ (0.477 g, 1.81 mmol)at 0° C. & then stirred for 5 min. After that CBr₄ (0.636 g, 1.92 mmol)was added & stirred at room temperature for 1 hr. After completion ofreaction, turbid reaction mass was filtered & washed with small amountof DCM to afford the desired product as white solid (0.27 g, 52.9%).

Step 5: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-(4-(bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in dry DMF, morpholine (5 equiv.) was added at 0° C. andreaction mixture was stirred for overnight. After completion ofreaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water; dried overanhydrous Na₂SO₄; filtered and concentrated under reduced pressure. Theresidue was purified by column chromatography/preparative HPLC to givethe desired compound (15% yield). LCMS: 515.25 (M+1)⁺; HPLC: 99.80% (@254 nm) (R_(t); 5.168); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 10.01(bs, 1H), 8.94 (bs, 1H), 8.39 (s, 1H), 8.35 (d, J=7.6 Hz, 2H), 8.18 (s,1H), 7.68 (d, J=7.2 Hz, 2H), 5.90 (s, 1H), 5.30 (m, 1H), 4.44-4.41 (m,4H), 3.98 (m, 2H), 3.63-3.60 (m, 2H), 3.37-3.30 (m, 2H), 3.16 (m, 2H),2.23 (s, 3H), 2.13 (s, 3H), 1.55 (d, J=6.4 Hz, 6H).

Synthesis of Compound A-3:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-((dimethylamino)methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A solution of ethyl1-isopropyl-6-(((trifluoromethyl)sulfonyl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(1 g, 2.62 mmol), 3-(hydroxymethyl) phenyl boronic acid (0.438 g, 2.88mmol) and Pd(PPh₃)₄ (0.303 g, 0.262 mmol) in 1,4-dioxane (15 mL) waspurged with argon for 10 min. Then 2M Na₂CO₃ solution (4.5 mL, 9.4 mmol)was added to it and again argon was purged through it for 10 min. Thereaction mixture was stirred at 100° C. for 1 h and after completion ofthe reaction water was added to it. Extraction was carried out usingEtOAc the combined organic layers were washed with water dried overanhydrous Na₂SO₄; filtered and concentrated under reduced pressure. Theresidue was purified by column chromatography to give 0.81 g of targetcompound.

Step 2: Synthesis of6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid

To a solution of ethyl6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.81 g, 2.38 mmol) in EtOH (15 mL) aqueous NaOH (0.47 g, 11.94 mmol)was added and stirred at 60° C. for 1 h. After completion of thereaction, ethanol was removed under reduced pressure and it wasacidified using 10% citric acid solution till pH 4. Extraction wascarried out using 5% MeOH/DCM, the combined organic layers were washedwith water & dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to afford the desired acid which was used withoutfurther purification.

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A mixture of6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid (0.65 g, 2.09 mmol), 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one(0.64 g, 4.18 mmol) and PYBOP (1.63 g, 3.13 mmol) in DMSO (5 mL) wasstirred at room temperature for 12 hr. After completion of the reaction,saturated NaHCO₃ solution was added to it and extraction was carried outusing 20% MeOH/DCM. The combined organic layers were washed with water;dried over anhydrous Na₂SO₄; filtered and concentrated under reducedpressure. The residue was purified by column chromatography to give awhite solid (0.71 g, 81.4%).

Step 4: Synthesis of6-(3-(bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide (0.45 g,0.1.011 mmol) in a DCM (10 mL), PPh₃(0.477 g, 1.81 mmol) was added at 0°C. & then stirred for 5 min. After that CBr₄ (0.636 g, 1.92 mmol) wasadded and stirred at room temperature for 1 hr. After completion ofreaction, solid was filtered & washed with small amount of DCM to afford0.30 g of pure desired product.

Step 5: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-((dimethylamino)methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-(3-(bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in a DMF, dimethylamine (5 equiv.) was added at 0° C. &resulting reaction mixture stirred for overnight. After completion ofreaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toobtain the crude material which was purified by column chromatography togive pure final compounds (40% yield). LCMS: 473.20 (M+1)⁺; HPLC: 99.89%(@ 254 nm) (R_(t); 5.325); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H),8.96 (bs, 1H), 8.35 (s, 1H), 8.13-8.10 (m, 3H), 7.50 (t, J=8.4 Hz, 1H),7.43-7.41 (m, 1H), 5.89 (s, 1H), 5.34-5.31 (m, 1H), 4.41 (d, J=4 Hz,2H), 3.50 (s, 2H), 2.23 (s, 3H), 2.19 (s, 6H), 2.12 (s, 3H), 1.54 (d,J=6.8 Hz, 6H).

Synthesis of Compound A-4:6-(2-aminopyrimidin-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-bromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

To a stirred solution of ethyl6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (20 g, 74.3 mmol) inacetonitrile (200 mL), K₂CO₃ (15.39 g, 111.52 mmol) and 2-bromopropane(18.13 g, 148.64 mmol) was added to it. The reaction mixture wasrefluxed for 8 h. On completion, acetonitrile was removed under reducedpressure and water was added to it. Extraction was carried out usingethyl acetate; the combined organic layers were washed with water, brineand dried over anhydrous Na₂SO₄. Solvent was removed under reducedpressure and residue was purified by silica gel column chromatography toobtain the desired intermediate (13.5 g, 58.3%).

Step 2: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (0.964 g, 24.11 mmol) was added to a solution of ethyl6-bromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (5 g, 16.07mmol) in EtOH (50 mL) and stirred at 60° C. for 1 h. After completion ofthe reaction, ethanol was removed under reduced pressure and it wasacidified using 10% citric acid solution. Extraction was carried outusing ethyl acetate; the combined organic layers were washed with water,brine and dried over anhydrous Na₂SO₄; filtered and concentrated underreduced pressure. The crude acid (3 g, 10.55 mmol) was then dissolved inDMSO (35 mL) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (3.2 g,21.0 mmol) was added to it. The reaction mixture was stirred at roomtemperature for 15 min before PYBOP (8.23 g, 15.82 mmol) was added to itand stirring was continued overnight. After completion of the reaction,reaction mass was poured into ice to obtain solid, which was filteredand washed with acetonitrile followed by ether to provide the desiredintermediate (3 g, 68%).

Step 3: Synthesis of6-(2-aminopyrimidin-5-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.), (2-aminopyrimidin-5-yl)boronic acid (1.2 equiv.) andPd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged with argon for 10 min.Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to it and again argonwas purged through it for 10 min. The reaction mixture was stirred at100° C. for 1 h. After completion of the reaction, water was added to itand extraction was carried out using EtOAc. The combined organic layerswere washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give the desired compound (45%yield). LCMS: 433.20 (M+1)⁺; HPLC: 98.75% (@ 254 nm) (R_(t); 5.252); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 9.09 (s, 2H), 8.84 (t, 1H), 8.33(s, 1H), 8.07 (s, 1H), 7.15 (s, 2H), 5.89 (s, 1H), 5.32-5.28 (m, 1H),4.40 (d, 2H, J=4.0 Hz), 2.22 (s, 3H), 2.12 (s, 3H), 1.51 (d, 6H, J=6.8Hz).

Synthesis of Compound A-5:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(2-morpholinoethyl)-1H-pyrazol-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.), (1-(2-morpholinoethyl)-1H-pyrazol-4-yl)boronic acid (1.2equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged with argonfor 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to it andagain argon was purged through it for 10 min. The reaction mixture wasstirred at 100° C. for 1 h. After completion of the reaction, water wasadded to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound. LCMS: 519.00 (M+1)⁺; HPLC: 97.07% (@ 254 nm) (R_(t); 4.925;Method: Column: YMC ODS-A 150 mm×4.6 mm×5μ Mobile Phase: A; 0.05% TFA inwater/B; 0.05% TFA in acetonitrile; Inj. Vol: 10 μL, Col. Temp.: 30° C.;Flow rate: 1.4 mL/min.; Gradient: 5% B to 95% B in 8 min, Hold for 1.5min, 9.51-12 min 5% B); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 8.76(t, 1H), 8.46 (s, 1H), 8.26 (s, 1H), 8.14 (s, 1H), 7.85 (s, 1H), 5.89(s, 1H), 5.34-5.31 (m, 1H), 4.39 (d, 2H, J=4.8 Hz), 4.30 (t, 2H, J=6.8Hz), 3.54 (m, 4H), 2.76 (t, 2H, J=6.4 Hz), 2.42 (m, 4H), 2.22 (s, 3H),2.12 (s, 3H), 1.51 (d, 6H, J=6.4 Hz).

Synthesis of Compound A-6: tert-butyl4-(5-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridin-6-yl)pyridin-2-yl)piperazine-1-carboxylate

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) tert-butyl4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)piperazine-1-carboxylate(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredproduct (70% yield). LCMS: 601.30 (M+1)⁺; HPLC: 99.80% (@ 254 nm),(R_(t); 6.124); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.54 (s, 1H), 9.03 (d, 1H,J=2 Hz), 8.85 (t, 1H), 8.39 (dd, 1H, J=8.8, 2 Hz), 8.31 (s, 1H), 8.08(s, 1H), 6.99 (d, 1H, J=9.2 Hz), 5.89 (s, 1H), 5.32-5.25 (m, 1H), 4.40(d, 2H, J=4.4 Hz), 3.63 (t, 4H), 3.45 (bs, 4H), 2.22 (s, 3H), 2.12 (s,3H), 1.53 (d, 6H, J=6.4 Hz), 1.43 (s, 9H).

Synthesis of Compound A-7:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(piperazin-1-yl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv), tert-butyl4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine-1-carboxylate(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the Bocprotected intermediate. Boc-deprotection was achieved by using TFA-DCM(10 times by volume in 1:1 ratio, work up using NaHCO₃) to give thedesired compound (65% yield). LCMS: 500.15 (M+1)⁺; HPLC: 99.23% (@ 254nm), (R_(t); 5.242); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.60 (bs, 1H), 9.23(bs, 2H), 8.94 (bs, 1H), 8.31 (s, 1H), 8.19 (d, 2H, J=8.8 Hz), 8.08 (s,1H), 7.14 (d, 2H, J=8.8 Hz), 5.91 (s, 1H), 5.33-5.26 (m, 1H), 4.40 (d,2H, J=4.8 Hz), 3.51 (bs, 4H), 3.24 (bs, 4H), 2.22 (s, 3H), 2.13 (s, 3H),1.53 (d, 6H, J=6.4 Hz).

Synthesis of Compound A-8:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-(4-(bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in dry DMF, 1-methylpiperazine (5 equiv.) was added at 0° C.and reaction mixture was stirred for overnight. After completion ofreaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water; dried overanhydrous Na₂SO₄; filtered and concentrated under reduced pressure. Theresidue was purified by column chromatography/preparative HPLC to givethe desired compound (5% yield). LCMS: 528.25 (M+1)+; HPLC: 98.21% (@254 nm) (Rt; 4.802); 1H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 8.94(bs, 1H), 8.36 (s, 1H), 8.19 (d, J=8.4 Hz, 2H), 8.13 (s, 1H), 7.46 (d,J=7.6 Hz, 2H), 5.89 (s, 1H), 5.33-5.30 (m, 1H), 4.41 (d, J=4 Hz, 2H),3.52 (s, 3H), 2.30-2.45 (m, 7H), 2.21 (s, 3H), 2.15 (s, 3H), 2.12 (s,3H), 1.54 (d, J=6.8 Hz, 6H).

Synthesis of Compound A-9:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(4-methylpiperazin-1-yl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv),1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound (57%). LCMS: 514.20 (M+1)+; HPLC: 99.74% (@ 254 nm), (Rt;5.183); 1H NMR (CDCl3, 400 MHz) δ 11.88 (bs, 1H), 8.30 (s, 1H), 8.16 (t,1H), 8.08 (d, 2H, J=8.8 Hz), 7.91 (s, 1H), 6.97 (d, 2H, J=8.8 Hz), 5.94(s, 1H), 5.43-5.36 (m, 1H), 4.66 (d, 2H, J=5.6 Hz), 3.30 (bs, 4H), 2.58(bs, 4H), 2.41 (s, 3H), 2.36 (s, 3H), 2.19 (s, 3H), 1.60 (d, 6H, J=6.4Hz).

Synthesis of Compound A-10:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(6-(piperazin-1-yl)pyridin-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv), tert-butyl4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)piperazine-1-carboxylate(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the Bocprotected intermediate. Boc-deprotection was achieved by using TFA-DCM(10 times by volume in 1:1 ratio, work up using NaHCO₃) to give thedesired compound (50% yield). LCMS: 501.20 (M+1)+; HPLC: 98.58% (@ 254nm), (Rt; 4.728); 1H NMR (DMSO-d6, 400 MHz) δ 11.56 (s, 1H), 9.07 (d,1H, J=2 Hz), 8.87 (t, 1H), 8.82 (bs, 2H), 8.46 (dd, 1H, J=8.8, 2.4 Hz),8.33 (s, 1H), 8.11 (s, 1H), 7.09 (d, 1H, J=8.8 Hz), 5.91 (s, 1H),5.33-5.26 (m, 1H), 4.40 (d, 2H, J=4.8 Hz), 3.84 (t, 4H), 3.24 (bs, 4H),2.23 (s, 3H), 2.13 (s, 3H), 1.54 (d, 6H, J=6.8 Hz).

Synthesis of Compound A-11:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(6-(4-methylpiperazin-1-yl)pyridin-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv),1-methyl-4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-yl)piperazine(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give the desiredcompound (30% yield). LCMS: 515.20 (M+1)+; HPLC: 98.36% (@ 254 nm), (Rt;4.876); 1H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 9.01 (s, 1H), 8.85(t, 1H), 8.36 (d, 1H, J=8.8 Hz), 8.31 (s, 1H), 8.07 (s, 1H), 6.97 (d,1H, J=8.4 Hz), 5.89 (s, 1H), 5.32-5.25 (m, 1H), 4.40 (d, 2H, J=4.4 Hz),3.61 (t, 4H), 2.41 (t, 4H), 2.22 (s, 6H), 2.12 (s, 3H), 1.53 (d, 6H,J=6.8 Hz).

Synthesis of Compound A-12:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(4-methylpiperazine-1-carbonyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv),(4-methylpiperazin-1-yl)(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give the desiredcompound (33% yield). LCMS: 542.20 (M+1)+; HPLC: 98.72% (@ 254 nm), (Rt;5.034); 1H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 8.96 (bs, 1H), 8.39(s, 1H), 8.32 (d, 2H, J=8 Hz), 8.20 (s, 1H), 7.56 (d, 2H, J=7.6 Hz),5.90 (s, 1H), 5.35-5.32 (m, 1H), 4.42 (d, 2H, J=4 Hz), 3.64 (bs, 2H),3.38 (m, 2H), 2.37 (bs, 2H), 2.28 (bs, 2H), 2.22 (s, 3H), 2.20 (s, 3H),2.12 (s, 3H), 1.55 (d, 6H, J=6.4 Hz).

Synthesis of CompoundA-13:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-((3-(dimethylamino)propyl)carbamoyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv),N-(3-(dimethylamino)propyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give the desiredcompound (9% yield). LCMS: 544.20 (M+1)+; HPLC: 98.83% (@ 254 nm), (Rt;5.258); 1H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 9.00 (t, 1H), 8.70(t, 1H), 8.64 (s, 1H), 8.39 (s, 1H), 8.38 (s, 1H), 8.19 (s, 1H), 7.92(d, 1H, J=8 Hz), 7.64 (t, 1H, J=7.6 Hz), 5.90 (s, 1H), 5.37-5.34 (m,1H), 4.41 (d, 2H, J=4.4 Hz), 2.28 (t, 2H, J=6.8 Hz), 2.23 (s, 3H), 2.14(s, 6H), 2.12 (s, 3H), 1.69 (t, 2H, J=6.8 Hz), 1.55 (d, 6H, J=6.4 Hz).

Synthesis of Compound A-14:6-(4-(aminomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzylcarbamate (1.2equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged with argonfor 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to it andagain argon was purged through it for 10 min. The reaction mixture wasstirred at 100° C. for 1 h. After completion of the reaction, water wasadded to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the Bocprotected intermediate. Boc-deprotection was achieved by using TFA-DCM(10 times by volume in 1:1 ratio, work up using NaHCO₃) to give thedesired compound (65% yield). LCMS: 445.15 (M+1)+; HPLC: 99.78% (@ 254nm), (Rt; 4.950); 1H NMR (DMSO-d6, 400 MHz) δ 11.52 (s, 1H), 8.91 (t,1H), 8.36 (s, 1H), 8.31 (d, 2H, J=8 Hz), 8.21 (bs, 2H), 8.16 (s, 1H),7.61 (d, 2H, J=8.8 Hz), 5.88 (s, 1H), 5.34-5.30 (m, 1H), 4.40 (d, 2H,J=4.4 Hz), 4.12 (d, 2H, J=5.2 Hz), 2.21 (s, 3H), 2.11 (s, 3H), 1.53 (d,6H, J=6.8 Hz).

Synthesis of Compound A-15:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(((3-(dimethylamino)propyl)(methyl)amino)methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(hydroxyl-methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(2.5 g, 5.98 mmol), (4-(hydroxymethyl)phenyl)boronic acid (1 g, 6.57mmol) and Pd(PPh₃)₄ (0.69 g, 0.597 mmol) in 1,4-dioxane (30 mL) waspurged with argon for 10 min. Then, 2 M Na₂CO₃ (2.3 g, 21.69 mmol) inwater was added to it and again argon was purged through it for 10 min.The reaction mixture was stirred at 100° C. for 2 h. After completion ofthe reaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound (2.6 g, 96.3%).

Step 2: Synthesis of6-(4-(bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(hydroxyl-methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(2.6 g, 5.84 mmol) in DCM (50 mL), triphenyl phosphine (6.1 g, 23.23mmol) was added and stirred it at room temperature for 10 min. FinallyCBr₄ (7.7 g, 23.23 mmol) was added portion-wise to it and resultingsolution was stirred at room temperature for overnight. After completionof reaction, solid was filtered and dried to obtain the desired compound(1 g, 34.5%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(((3-(dimethylamino)propyl)(methyl)amino)methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

6-(4-(Bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was suspended in DMF andN1,N1,N3-trimethylpropane-1,3-diamine (5 equiv.) was added to it. Thereaction mixture was stirred at room temperature for overnight. Oncompletion of reaction, solvent was removed under reduced pressure andresidue was purified by prep. HPLC chromatography to provide the desiredcompound (30% yield). LCMS: 544.25 (M+1)⁺; HPLC: 99.96% (@ 254 nm)(R_(t); 4.732); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (bs, 1H), 10.14-9.80(m, 1H), 8.95 (bs, 1H), 8.40-8.35 (m, 3H), 8.22-8.19 (m, 1H), 7.75-7.69(m, 2H), 5.91 (s, 1H), 5.35-5.32 (m, 1H), 4.42 (d, 2H, J=3.6 Hz),3.08-2.98 (m, 6H), 2.80-2.62 (m, 8H), 2.23 (s, 3H), 2.13 (s, 3H), 2.07(m, 2H). 1.55 (d, 6H, J=6.4 Hz).

Synthesis of Compound A-16:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(((3-(dimethylamino)propyl)(methyl)amino)methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(2.5 g, 5.98 mmol), (3-(hydroxymethyl)phenyl)boronic acid (1 g, 6.57mmol) and Pd(PPh₃)₄ (0.69 g, 0.598 mmol.) in 1,4-dioxane (30 mL) waspurged with argon for 10 min. Then, 2 M Na₂CO₃(2.3 g, 21.69 mmol) inwater was added to it and again argon was purged through it for 10 min.The reaction mixture was stirred at 100° C. for 2 h. After completion ofthe reaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound (2.1 g, 80.7%).

Step 2: Synthesis of6-(3-(bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(2.1 g, 4.7 mmol) in DCM (35 mL), triphenyl phosphine (5 g, 19.06 mmol)was added and stirred it at room temperature for 10 min. Finally CBr₄(6.2 g, 18.69 mmol) was added portion wise to it and resulting solutionwas stirred at room temperature for overnight. After completion ofreaction, solid was filtered and dried to obtain the desired compound(1.2 g, 52.2%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(((3-(dimethylamino)propyl)(methyl)amino)methyl)phenyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

6-(3-(Bromomethyl)phenyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was suspended in DMF andN1,N1,N3-trimethylpropane-1,3-diamine (5 equiv.) was added to it. Thereaction mixture was stirred at room temperature for overnight. Oncompletion of reaction, solvent was removed under reduced pressure andthe residue was purified by prep. HPLC chromatography to provide thedesired compound (25% yield). LCMS: 544.30 (M+1)⁺; HPLC: 99.90% (@ 254nm) (R_(t); 4.765); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (bs, 1H), 10.2-9.6(m, 1H), 8.92 (bs, 1H), 8.38 (s, 2H), 8.14 (s, 1H), 7.70-7.66 (m, 2H),5.91 (s, 1H), 5.36-5.35 (m, 1H), 4.41 (d, 2H), 3.39 (4H merged in DMSOpeak), 3.08-2.98 (m, 2H+3H), 2.79 (s, 6H), 2.24 (s, 3H), 2.13 (s, 3H).2.13 (m, 2H), 1.55 (d, 6H, J=4.8 Hz).

Synthesis of Compound A-17:3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl 3,6-dibromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

To a stirred solution of ethyl6-bromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (2 g, 6.43mmol) in acetic acid (6 mL) was added bromine(5.14 g, 32.15 mmol) at 0°C. Resulting reaction mass was stirred at room temperature for 3 h.Reaction was monitored by TLC. On completion, reaction was quenched withice, NaHCO₃ solution was added to it. Extraction was carried out usingethyl acetate; the combined organic layers were washed with water, brineand dried over anhydrous Na₂SO₄. Solvent was removed under reducedpressure and residue was purified by silica gel column chromatography toobtain the desired intermediate (1.2 g, 48%).

Step 2: Synthesis of ethyl3-bromo-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A solution of ethyl 3,6-dibromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1.2 g,3.08 mmol),4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine(0.935 g, 3.08 mmol) and Pd(PPh₃)₄ (0.178 g, 0.154 mmol) in 1,4-dioxane(33 mL) was purged with argon for 10 min. Then, 2M Na₂CO₃ solution (1.17g, 11.10 mmol) was added to it and again argon was purged through it for10 min. The reaction mixture was stirred at 100° C. for 1 h. Aftercompletion of the reaction, water was added to it and extraction wascarried out using 10% MeOH/DCM. The combined organic layers were washedwith water, dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to afford crude material which was purified by columnchromatography to give the desired compound (0.8 g, 67.1%).

Step 3: Synthesis of3-bromo-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid

To a solution of ethyl3-bromo-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(1 g, 2.05 mmol) in EtOH (15 mL), aqueous NaOH (0.123 g, 3.08 mmol) wasadded and reaction mixture stirred at 60° C. for 1 h. After completionof the reaction, ethanol was removed under reduced pressure and it wasacidified using 10% citric acid solution. Solid obtained was filteredand azeotrope it with toluene to give the desired compound (0.8 g,84.9%).

Step 4: Synthesis of3-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of3-bromo-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid (0.8 g, 1.74 mmol) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.531 g, 3.49 mmol) inDMSO (8 mL) and was stirred at room temperature for 15 min. Then PYBOP(1.36 g, 2.62 mmol) was added to it and stirring was continued for 12hr. After completion of the reaction, reaction mass was poured into iceto obtain solid, which was filtered and solid was purified by silica gelcolumn chromatography to obtain the desired compound (39% yield). LCMS:593.20 (M+1)⁺; HPLC: 97.49% (@ 254 nm) (R_(t); 5.338); ¹H NMR (DMSO-d₆,400 MHz) δ 11.48 (bs, 1H), 8.67 (t, 1H, J=4.8 Hz), 8.18 (d, 2H, J=8.4Hz), 7.73 (s, 1H), 7.47 (d, 2H, J=8.4 Hz), 5.87 (s, 1H), 5.33-5.30 (m,1H), 4.40 (d, 2H, J=4.4 Hz), 3.58 (bs, 4H), 3.53 (s, 2H), 2.38 (bs, 4H),2.26 (s, 3H), 2.11 (s, 3H), 1.52 (d, 6H, J=6.4 Hz).

Synthesis of Compounds A-18 through A-78, A-91 through A-110, A-125, andA-126

Compounds A-18 through A-78, A-91 through A-110, A-125, and A-126 weresynthesized by methods similar to those described for Compounds A-1through A-17 or by reaction schemes depicted in the general schemes.

Synthesis of Compound B-1:1-(1-benzylpiperidin-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one

3-Cyano-2,4-dimethyl-2-hydroxypyridine (0.3 g, 2.0 mmol) was dissolvedin MeOH (5 mL), to which catalytic amount of Raney Ni and of aqueous NH₃(0.3 mL) were added and the reaction mixture was stirred under hydrogenpressure (bladder pressure) for 3-4 h. After completion of the reaction,catalyst was filtered off and the filtrate was concentrated underreduced pressure. The residue was thoroughly dried to provide thedesired product (0.3 g, quantitative yield).

Step 2: Synthesis of ethyl6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A stirred solution of 1H-pyrazol-3-amine (45 g, 542 mmol) in acetic acid(297 mL) and water (900 mL) was cooled to 0° C. and diethyl oxaloacetatesodium salt (113.85 g, 542.16 mmol) was added to it. Resulting solutionwas heated at 100° C. for overnight. After completion of reaction, solidwas filtered and dried to obtain the desired intermediate (25 g, 22%).

Step 3: Synthesis of ethyl6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (25 g, 120mmol) was suspended in acetonitrile (250 mL) and POBr₃ (69.56 g, 241.54mmol) was added to it. The reaction mixture was refluxed for 6 h. Oncompletion of reaction, acetonitrile was removed under reduced pressureand residue neutralized with saturated NaHCO₃ solution was added to it.Extraction was carried out using ethyl acetate; the combined organiclayers were washed with water, brine and dried over anhydrous Na₂SO₄;filtered and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography to provide the desiredintermediate (25 g, 77%).

Step 4: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (1.11 g, 27.88 mmol) was added to a solution of ethyl6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (5 g, 18.58 mmol) inEtOH (50 mL) and stirred at 60° C. for 1 h. After completion of thereaction, ethanol was removed under reduced pressure and it wasacidified using 10% citric acid solution. Solid obtained was filteredand dried under reduced pressure to obtain corresponding acid. The crudeacid (4.4 g, 18.2 mmol) was then dissolved in DMSO (20 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (5.55 g, 36.51 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (14.24 g, 27.38 mmol) was added to it and stirring wascontinued overnight. After completion of the reaction, reaction mass waspoured into ice to obtain a solid, which was filtered and washed withacetonitrile followed by ether to provide the desired intermediate (4 g,58.5%).

Step 5: Synthesis of tert-butyl4-(6-bromo-4-4(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-pyrazolo[3,4-b]pyridin-1-yl)piperidine-1-carboxylate

To a stirred solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.5 g, 1.33 mmol) in DMF (5 mL), K₂CO₃ (0.275 g, 1.99 mmol) andtert-butyl 4-bromopiperidine-1-carboxylate (0.529 g, 2.01 mmol) wasadded. Resulting reaction mixture was stirred at 80° C. for 12 h. Oncompletion of reaction, water was added to it and extraction was carriedout using DCM; the combined organic layers were washed with water, brineand dried over anhydrous Na₂SO₄. Solvent was removed under reducedpressure and residue was purified by silica gel column chromatography toobtain the desired intermediate (0.5 g, 67.2%).

Step 6: Synthesis of tert-butyl4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)piperidine-1-carboxylate

A solution of tert-butyl4-(6-bromo-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-pyrazolo[3,4-b]pyridin-1-yl)piperidine-1-carboxylate(0.5 g, 0.896 mmol), boronic ester (0.285 g, 1.075 mmol) and Pd(PPh₃)₄(0.103 g, 0.089 mmol) in 1,4-dioxane (8 mL) was purged with argon for 10min. Then, 2M Na₂CO₃ (0.341 g, 3.216 mmol) was added to it and argon waspurged again for 10 min. The reaction mixture was stirred at 100° C. for1 h. After completion of the reaction, water was added to it andextraction was carried out using 10% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by prep HPLC to give the desired compound (0.5 g, 90.5%).

Step 7: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of tert-butyl4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridin-1-yl)piperidine-1-carboxylate(0.5 g, 0.8 mmol) in DCM (10 mL), TFA (0.3 mL) was added at 0° C. andreaction stirred at room temperature for 1 h. After completion ofreaction, solvent was removed under reduced pressure and residueneutralized with saturated NaHCO₃ solution Extraction was carried outusing DCM; the combined organic layers were washed with water, brine anddried over anhydrous Na₂SO₄; filtered and concentrated under reducedpressure. Finally the residue was washed with diethyl ether to providethe desired compound (0.4 g, 95.7%).

Step 8: Synthesis of1-(1-benzylpiperidin-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.1 g, 0.193 mmol) in DMF (1 mL), K₂CO₃(0.032 g, 0.232 mmol) and benzylbromide (0.04 g, 0.232 mmol) were added. Resulting reaction mixture wasstirred at 80° C. for 12 h. On completion of reaction, water was addedto it and extraction was carried out using DCM; the combined organiclayers were washed with water, brine and dried over anhydrous Na₂SO₄.Solvent was removed under reduced pressure and residue was purified byprep HPLC to provide the desired compound (21.3% yield). LCMS: 608.45(M+1)⁺; HPLC: 94.92% (@ 254 nm) (R_(t); 4.640); ¹H NMR (DMSO-d₆, 400MHz) δ 11.54 (s, 1H), 9.70 (bs, 1H), 8.86 (s, 1H), 8.37 (s, 1H), 7.90(s, 1H), 7.54-7.52 (m, 5H), 6.85 (s, 1H), 5.90 (s, 1H), 5.17 (m, 1H),4.40 (m, 4H), 3.55 (d, 2H, J=12 Hz), 3.41-3.35 (m, 2H), 2.83 (m, 2H),2.32-2.41 (m, 2H), 2.23 (s, 3H), 2.22-2.13 (m, 2H), 2.12 (s, 3H), 1.57(s, 6H), 1.47 (s, 6H).

Synthesis of Compound B-2:1-(1-acetylpiperidin-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.1 g, 0.2 mmol) was suspended in pyridine (1 mL) and acetyl chloride(0.016 g, 0.205 mmol) was added to it. The reaction mixture was stirredat room temperature for 5 h. On completion of reaction, water was addedto it and extraction was carried out using 10% MeOH/DCM; the combinedorganic layers were washed with water, brine and dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure. The residuewas purified by prep HPLC to provide the desired compound (30% yield).LCMS: 560.00 (M+1)⁺; HPLC: 99.09% (@ 254 nm) (R_(t); 4.685); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.56 (s, 1H), 8.85 (t, 1H, J=5.2 Hz), 8.74 (s,2H), 8.32 (s, 1H), 7.87 (s, 1H), 6.84 (s, 1H), 5.90 (s, 1H), 5.12-5.06(m, 1H), 4.48 (d, 2H, J=12 Hz), 4.38 (d, 2H, J=5.2 Hz), 3.97 (m, 2H),3.33-3.30(m, 1H), 2.83 (s, 2H), 2.23 (s, 3H), 2.12 (s, 3H), 2.08 (s,3H), 2.03-1.91 (m, 4H), 1.57 (s, 6H) 1.47 (s, 6H).

Synthesis of Compound B-3:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(2-methylallyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(2-methylallyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.4 g, 1.0 mmol) in DMF (4 mL), K₂CO₃ (0.152 g, 1.10 mmol) and 3chloro-2-methylpropene (0.115 g, 1.27 mmol) were added. Resultingreaction mixture was stirred at 80° C. for 2 h. On completion ofreaction, water was added to it and extraction was carried out using 20%MeOH/DCM; the combined organic layers were washed with water, brine anddried over anhydrous Na₂SO₄. Solvent was removed under reduced pressureand residue was purified by silica gel column chromatography to obtainthe desired intermediate (0.2 g, 43.8%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(2-methylallyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(2-methylallyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.2 g, 0.45 mmol), boronic ester (0.147 g, 0.554 mmol) and Pd(PPh₃)₄(0.053 g, 0.045 mmol) in 1,4-dioxane (3 mL) was purged with argon for 10min. Then, 2M Na₂CO₃ solution (0.175 g, 1.65 mmol) was added to it andargon was purged again for 10 min. The reaction mixture was stirred at100° C. for 1 h. After completion of the reaction, water was added to itand extraction was carried out using 10% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give the desired compound (44.2%yield). LCMS: 489.30 (M+1)⁺; HPLC: 97.19% (@ 254 nm) (R_(t); 5.112); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 8.90 (t, 1H), 8.29 (s, 1H), 7.81(s, 1H), 6.81 (s, 1H), 5.89 (s, 1H), 5.02 (s, 2H), 4.87 (s, 1H), 4.66(s, 1H), 4.38 (d, 2H, J=5.2 Hz), 2.42 (m, 2H), 2.23 (s, 3H), 2.12 (s,3H), 1.61 (s, 3H), 1.24 (s, 6H), 1.14 (s, 6H).

Synthesis of Compounds B-4 and B-5:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethylpiperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamideand1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-bromo-1-cyclopentyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (3 g, 11.15 mmol)was dissolved in acetonitrile (30 mL) and K₂CO₃ (1.85 g, 13.38 mmol) andcyclopentyl bromide (3.35 g, 22.30 mmol) was added to it. The reactionmixture was refluxed for 3 h. On completion, acetonitrile was removedunder reduced pressure and water was added to it. Extraction was carriedout using ethyl acetate; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄. Solvent was removed underreduced pressure and residue was purified by silica gel columnchromatography to obtain the desired intermediate (1.3 g, 50%).

Step 2: Synthesis of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (0.308 g, 7.72 mmol) was added to a solution of ethyl6-bromo-1-cyclopentyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1.3 g,3.8 mmol) in EtOH (10 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand it was acidified using 10% citric acid solution. Extraction wascarried out using ethyl acetate; the combined organic layers were washedwith water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure. The crude acid (1 g, 3.23 mmol) wasthen dissolved in DMSO (10 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.98 g, 6.47 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (2.52 g, 4.85 mmol) was added to it and stirring wascontinued overnight. After completion of the reaction, reaction mass waspoured into ice to obtain solid, which was filtered and washed withacetonitrile followed by ether to provide the desired intermediate (1 g,70%).

Step 3: Synthesis of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(Compound B-5)

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.5 g, 1.128 mmol), boronic ester (0.28 g, 1.24 mmol) and Pd(PPh₃)₄(0.13 g, 0.11 mmol) in 1,4-dioxane (5 mL) was purged with argon for 10min. Then, 2 M Na₂CO₃ (0.43 g, 4.063 mmol) in water was added to it andagain argon was purged through it for 10 min. The reaction mixture wasstirred at 100° C. for 1 h. After completion of the reaction, water wasadded to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound (0.25 g, 44.1%). LCMS: 503.30 (M+1)⁺; HPLC: 97.176% (@ 254 nm)(R_(t); 5.599); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (bs, 1H); 8.86 (s, 1H); 8.28 (s, 1H); 7.81 (s, 1H); 6.82 (s, 1H); 5.89 (s, 1H); 5.42-5.38(m, 1H); 4.38 (d, J=4.8 Hz, 2H); 2.23 (s, 3H); 2.12 (s, 3H); 2.02-1.91(m, 6H); 1.72 (m, 2H); 1.38-1.23 (m, 14H).

Step 4: Synthesis of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethylpiperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.06 g, 0.119 mmol) in EtOH (10 mL), 10% Pd/C in catalytic amount wasadded and stirred it at room temperature under hydrogen pressure(bladder pressure) for 3-5 hr. On completion of reaction, reaction massfilter it through celite bed, then filtrate was concentrated underreduce pressure. The residue was washed with acetonitrile followed byether to provide the desired product (75% yield). LCMS: 505.30 (M+1)⁺;HPLC: 99.69% (@ 254 nm) (R_(t); 5.517); ¹H NMR (CD3OD-d₆, 400 MHz) δ11.54 (bs, 1H), 8.76 (t, 1H, J=4.8 Hz), 8.26 (s, 1H), 7.51 (s, 1H), 5.89(s, 1H), 5.38-5.36 (m, 1H), 4.36 (d, 2H, J=4.8 Hz), 2.21 (s, 3H), 2.12(s, 3H), 2.12 (m, 3H), 2.01-1.98 (m, 2H), 1.96-1.89 (m, 2H), 1.75-1.71(m, 4H), 1.52 (m, 2H), 1.27 (s, 6H), 1.12 (s, 6H).

Synthesis of Compound B-6:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-((1-methylpiperidin-4-yl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-bromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

To a stirred solution of ethyl6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (20 g, 74.3 mmol) inacetonitrile (200 mL), K₂CO₃ (15.39 g, 111.52 mmol) and 2-bromopropane(18.13 g, 148.64 mmol) was added to it. The reaction mixture wasrefluxed for 8 h. On completion, acetonitrile was removed under reducedpressure and water was added to it. Extraction was carried out usingethyl acetate; the combined organic layers were washed with water, brineand dried over anhydrous Na₂SO₄. Solvent was removed under reducedpressure and residue was purified by silica gel column chromatography toobtain the desired intermediate (13.5 g, 58.3%).

Step 2: Synthesis of ethyl1-isopropyl-6-((1-methylpiperidin-4-yl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-hydroxy-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1equiv.) was suspended in DMF and K₂CO₃ (1.5 equiv.) and4-bromo-1-methylpiperidine (1.5 equiv.) was added to it. The reactionmixture was stirred at room temperature for overnight. On completion ofreaction, water was added to it and extraction was carried out usingethyl acetate; the combined organic layers were washed with water, brineand dried over anhydrous Na₂SO₄. Solvent was removed under reducedpressure and residue was purified by silica gel column chromatography toobtain the desired intermediate.

Step 3: synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-((1-methylpiperidin-4-yl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (2 equivalent) was added to a solution of ethyl1-isopropyl-6-((1-methylpiperidin-4-yl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(1 equivalent) in EtOH (10 times by volume) and stirred at 60° C. for 1h. After completion of the reaction, ethanol was removed under reducedpressure and it was acidified using citric acid solution. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure. The residue was then dissolved in DMSO (10 timesby volume) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2equivalents) was added to it. The reaction mixture was stirred at roomtemperature for 15 min before PYBOP (1.5 equivalents) was added to itand stirring was continued for 5 to 16 h. After completion of thereaction, reaction mass was poured into ice water to obtain a solidwhich was filtered and washed with acetonitrile to provide the desiredproduct (2.5% yield). LCMS: 453.20 (M+1)⁺; HPLC: 96.37% (@ 254 nm)(R_(t); 4.742); ¹H NMR (CD3OD, 400 MHz) δ 8.16 (s, 1H), 7.21-7.11 (m,2H), 7.00 (s, 1H), 6.13 (s, 1H), 5.16-5.11 (m, 1H), 4.53 (d, 2H, J=5.2Hz), 3.65 (d, 1H, J=13.2 Hz), 3.48-3.45 (m, 2H), 3.38-3.31 (m, 2H), 2.94(s, 3H), 2.53 (d, 1H, J=12 Hz), 2.41 (s, 3H), 2.40 (m, 1H), 2.25 (s,3H), 2.20-2.13 (m, 1H), 2.02-1.95 (1H), 1.54 (d, 6H, J=6.8 Hz).

Synthesis of Compound B-7:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(4-(dimethylamino)butanoyl)piperidin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (0.964 g, 24.11 mmol) was added to a solution of ethyl6-bromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (5 g, 16.07mmol) in EtOH (50 mL) and stirred at 60° C. for 1 h. After completion ofthe reaction, ethanol was removed under reduced pressure and it wasacidified using 10% citric acid solution. Extraction was carried outusing ethyl acetate; the combined organic layers were washed with water,brine and dried over anhydrous Na₂SO₄; filtered and concentrated underreduced pressure. The crude acid (3 g, 10.55 mmol) was then dissolved inDMSO (35 mL) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (3.2 g,21.0 mmol) was added to it. The reaction mixture was stirred at roomtemperature for 15 min before PYBOP (8.23 g, 15.82 mmol) was added to itand stirring was continued overnight. After completion of the reaction,reaction mass was poured into ice to obtain solid, which was filteredand washed with acetonitrile followed by ether to provide the desiredintermediate (3 g, 68%).

Step 2: Synthesis of tert-butyl4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridin-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(3.1 g, 7.4 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate(2.7 g, 8.7 mmol) and Pd(PPh₃)₄ (0.428 g, 0.370 mmol.) in 1,4-dioxane(20 mL) was purged with argon for 10 min. Then, 2M Na₂CO₃ (2.8 g, 26.41)was added to it and again argon was purged through it for 10 min. Thereaction mixture was stirred at 100° C. for 1 h. After completion of thereaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired intermediate (2.8 g, 73.7%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of tert-butyl4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridin-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate(2.8 g, 5.4 mmol) in DCM (15 mL), TFA (13 mL) was added at 0° C. andstirred it at room temperature for 1 h. After completion of reaction,solvent was removed under reduced pressure and residue neutralized withsaturated NaHCO₃ solution Extraction was carried out using DCM; thecombined organic layers were washed with water, brine and dried overanhydrous Na₂SO₄; filtered and concentrated under reduced pressure,finally the residue was washed with diethyl ether to provide the desiredintermediate (1.5 g, 66.4%).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(4-(dimethylamino)butanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was dissolved in DMSO, to which Et₃N (3 equiv.) and4-(dimethylamino)butanoic acid (2 equiv.) was added. The reactionmixture was stirred at room temperature for 15 min before PYBOP (1.5equiv.) was added to it and stirring was continued overnight. Aftercompletion of the reaction, water was added to it. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to afford crude material which was purified bycolumn chromatography to give the desired product.

Step 5: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(4-(dimethylamino)butanoyl)piperidin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(4-(dimethylamino)butanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in EtOH, 10% Pd/C in catalytic amount was added and stirredit at room temperature under hydrogen pressure (bladder pressure) for3-16h hr. On completion of reaction, reaction mass was filtered throughcelite bed, then filtrate was concentrated under reduced pressure togive the desired product. LCMS: 536.40 (M+1)+; HPLC: 83.00% (@ 254 nm)(Rt:4.917); 1H NMR (DMSO-d6, 400 MHz) δ 11.54 (s, 1H), 8.75 (t, 1H),8.27 (s, 1H), 7.52 (s, 1H), 5.89 (s, 1H), 5.20-5.17 (m, 1H), 4.57-4.54(m, 1H), 4.36 (d, 2H, J=4.8 Hz), 4.01-3.98 (m, 1H), 3.32 (1H merged withDMSO peak), 3.19-305 (m, 2H), 2.72-2.67 (m, 2H), 2.40 (m, 2H), 2.40 (s,6H) 2.20 (s, 3H), 2.12 (s, 3H), 2.04-1.91 (m, 2H), 1.79-1.63 (m, 4H),1.48 (d, 6H, J=6.4 Hz).

Synthesis of Compound B-8:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylazetidine-3-carbonyl)piperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylazetidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was dissolved in DMSO, to which Et₃N (3 equiv.) and1-methylazetidine-3-carboxylic acid (2 equiv.) was added. The reactionmixture was stirred at room temperature for 15 min before PYBOP (1.5equiv.) was added to it and stirring was continued overnight. Aftercompletion of the reaction, water was added to it. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to afford crude material which was purified bycolumn chromatography to give the desired product.

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylazetidine-3-carbonyl)piperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylazetidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in EtOH, 10% Pd/C in catalytic amount was added and stirredit at room temperature under hydrogen pressure (bladder pressure) for3-16h hr. On completion of reaction, reaction mass was filtered throughcelite bed, then filtrate was concentrated under reduced pressure togive the desired product 62% yield). LCMS: 520.35 (M+1)⁺; HPLC: 93.45%(@ 254 nm) (R_(t); 6.931); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.54 (s, 1H),8.75 (t, 1H, J=4.8 Hz), 8.27 (s, 1H), 7.51 (s, 1H), 5.88 (s, 1H),5.20-5.17 (m, 1H), 4.50 (d, 1H, J=12.4 Hz), 4.36 (d, 2H, J=4.8 Hz), 3.69(d, 1H, J=13.2 Hz), 3.43 (s, 3H), 3.13-3.06 (m, 4H), 2.69 (t, 1H, J=12 &11.6 Hz), 2.20 (s, 3H), 2.17 (s, 3H), 2.12 (s, 3H), 1.93 (d, 2H, J=12Hz), 1.65 (m, 2H), 1.48 (d, 6H, J=7.2 Hz).

Synthesis of Compound B-9:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-methylpiperazin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1 Synthesis of ethyl1-isopropyl-6-(4-methylpiperazin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

To a stirred solution of ethyl6-bromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (0.3 g,0.964 mmol) in acetonitrile (10 mL), K₂CO₃ (0.26 g, 1.93 mmol) and1-methylpiperazine (0.289 g, 2.892 mmol) was added. The reaction mixturewas refluxed for 3 h. On completion of reaction, acetonitrile wasremoved under reduced pressure and water was added to it. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄. Solvent was removed underreduced pressure to obtain the desired intermediate (0.32 g, 94.9%).

Step 2: Synthesis of1-isopropyl-6-(4-methylpiperazin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid

Aqueous NaOH (0.193 g, 4.833 mmol) was added to a solution of ethyl1-isopropyl-6-(4-methylpiperazin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.32 g, 0.966 mmol) in EtOH (6 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand it was acidified using 10% citric acid solution. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine; dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to give the desired compound (0.2 g, 64.9%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-methylpiperazin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of1-isopropyl-6-(4-methylpiperazin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid (0.2 g, 0.600 mmol) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.182 g, 1.197 mmol) inDMSO (2 mL) and was stirred at room temperature for 15 min. Then PyBOP(0.468 g, 0.900 mmol) was added to it and stirring was continued for 12hr. After completion of the reaction, saturated NaHCO₃ solution wasadded to it and extraction was carried out using DCM. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by prep. HPLC to give the desired compound(53% yield). LCMS: 438.15 (M+1)⁺; HPLC: 96.67% (@ 254 nm), (R_(t);4.516); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.54 (s, 1H), 9.75 (s, 1H), 8.62(s, 1H), 8.06 (s, 1H), 7.20 (s, 1H), 5.89 (s, 1H), 5.04-4.99 (m, 1H),4.58 (d, 2H, J=14 Hz), 4.36 (d, 2H, J=4.4 Hz), 3.55 (d, 2H, J=11.20 Hz),3.24 (t, 2H, J=12.40 Hz), 3.12 (m, 2H), 2.84 (s, 3H), 2.20 (s, 3H), 2.12(s, 3H), 1.45 (d, 6H, J=6.8 Hz).

Synthesis of Compound B-10:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(tetrahydro-2H-pyran-4-yl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-bromo-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1 equiv.) wassuspended in acetonitrile and K₂CO₃ (1.5 equiv.) and4-bromotetrahydro-2H-pyran (2 equiv.) was added to it. The reactionmixture was refluxed for 8 h. On completion, acetonitrile was removedunder reduced pressure and water was added to it. Extraction was carriedout using ethyl acetate; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄. Solvent was removed underreduced pressure and residue was purified by silica gel columnchromatography to obtain the desired intermediate.

Step 2: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (5 equivalent) was added to a solution of ethyl6-bromo-1-cyclopentyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1equivalent) in EtOH (10 times by volume) and stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and it was acidified using 10% citric acid solution. Extractionwas carried out using EtOAc; the combined organic layers were washedwith water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure. The residue was then dissolved inDMSO (10 times by volume) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equivalents) was addedto it. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (1.5 equivalents) was added to it and stirring wascontinued for 5 to 16 h. After completion of the reaction, water wasadded to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water; brine; dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography followed by ether washto provide the desired intermediate.

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(tetrahydro-2H-pyran-4-yl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) solution was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound (14%). LCMS: 519.30 (M+1)⁺; HPLC: 98.464% (@ 254 nm) (R_(t);4.823); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (bs, 1H); 8.87 (s, 1 H); 8.30(s, 1H); 7.82 (s, 1H); 6.81 (s, 1H); 5.90 (s, 1H); 5.06 (m, 1H); 4.38(s, 2H); 3.01 (d, J=11.2, 2H); 3.59 (t, J=12.0 Hz, 2H); 2.23 (s, 3H);2.23-2.19 (m, 2H); 2.12 (s, 3H); 1.90 (d, J=12.0 Hz, 2H); 1.27-1.23 (m,14H).

Synthesis of Compound B-11:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-hydroxypiperidin-1-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-oxopiperidin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.1 g, 0.239 mmol) in DMF (2 mL), K₂CO₃ (0.198 g, 1.434 mmol) andpiperidin-4-one HCl salt (0.186 g, 0.94 mmol) were added. The reactionmixture was heated at 80° C. for 16 h. Reaction was monitored by LCMS &TLC. On completion, reaction mass was poured into water and extractionwas carried out using 5% MeOH/DCM. The combined organic layers weredried over anhydrous Na₂SO₄; filtered and concentrated under reducedpressure. The residue was purified by prep HPLC to provide the desiredcompound (0.03 g 28.8%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-hydroxypiperidin-1-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-oxopiperidin-1-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.1 g, 0.229 mmol) was taken in MeOH (3 mL) and NaBH₄ (0.020 g, 0.458mmol) was added at 0° C. The reaction mixture was stirred at roomtemperature for 4 h. On completion of reaction, MeOH was concentrated.And residue was purified by prep HPLC to provide the desired compound(0.034 g, 34%). LCMS: 439.20 (M+1)⁺; HPLC: 98.22% (@ 254 nm), (R_(t);5.393); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 8.63 (bs, 1H), 7.97(s, 1H), 7.11 (s, 1H), 5.88 (s, 1H), 4.98-4.95 (m, 1H), 4.34 (d, 2H,J=4.4 Hz), 4.15 (m, 2H), 3.74-3.72 (m, 1H), 3.23 (m, 3H), 2.19 (s, 3H),2.12 (s, 3H), 1.82 (m, 2H), 1.43 (d, 6H, J=6.4 Hz), 1.38 (m, 2H).

Synthesis of Compound B-12:5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1 Synthesis of ethyl5-chloro-6-hydroxy-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

To a stirred solution of ethyl6-hydroxy-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1.3 g,5.2 mmol) in DMF (7 mL), N-chlorosuccinamide (1.11 g, 8.35 mmol) wasadded. The resulting reaction mixture was stirred at 60° C. for 1 h. Oncompletion of reaction, water was added to it and extraction was carriedout using ethyl acetate; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄. Solvent was removed underreduced pressure and residue was purified by silica gel columnchromatography to obtain the desired intermediate (1 g, 69.9%).

Step 2 Synthesis of ethyl5-chloro-1-isopropyl-6-(((trifluoromethyl)sulfonyl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

To a stirred solution of ethyl5-chloro-6-hydroxy-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(1 g, 3.53 mmol) in DCM (10 mL), pyridine (0.336 mL, 4.24 mmol) wasadded followed by dropwise addition of triflic anhydride (0.716 mL, 4.24mmol). The reaction mixture was stirred at room temperature for 1 h. Oncompletion of reaction, water was added to it and extraction was carriedout using DCM; the combined organic layers were washed with water, brineand dried over anhydrous Na₂SO₄. Solvent was removed under reducedpressure and residue was purified by silica gel column chromatography toobtain the desired intermediate (0.8 g, 54.8%).

Step 3: Synthesis of ethyl5-chloro-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A solution of ethyl5-chloro-1-isopropyl-6-(((trifluoromethyl)sulfonyl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.2 g, 0.481 mmol),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(0.127 g, 0.481 mmol) and Pd(PPh₃)₄ (0.027 g, 0.024 mmol) in 1,4-dioxane(3 mL) was purged with argon for 10 min. Then, 2M Na₂CO₃ solution (0.183g, 1.734 mmol) was added to it and again argon was purged through it for10 min. The reaction mixture was stirred at 100° C. for 1 h. Aftercompletion of the reaction, water was added to it and extraction wascarried out using 10% MeOH/DCM. The combined organic layers were washedwith water, dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to afford crude material which was purified by columnchromatography to give the desired compound (0.09 g, 45.6%).

Step 4: Synthesis of5-chloro-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxyc acid

To a solution of intermediate 5 (0.09 g, 0.222 mmol) in EtOH (3 mL),aqueous NaOH (0.026 g, 0.668 mmol) was added and reaction mixturestirred at 60° C. for 1 h. After completion of the reaction, ethanol wasremoved under reduced pressure and it was acidified using 10% citricacid solution. Extraction was carried out using 5% MeOH/DCM. Thecombined organic layers were washed with water & dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford thedesired compound (0.07 g, 87.5%), which was pure enough for further use.

Step 5: Synthesis of5-chloro-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of5-chloro-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid (0.07 g, 0.18 mmol) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.056 g, 0.372 mmol) inDMSO (1 mL) and was stirred at room temperature for 15 min. Then PyBOP(0.145 g, 0.279 mmol) was added to it and stirring was continued for 12h. After completion of the reaction, water was added to it andextraction was carried out using 10% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by prep. HPLC to give the desired compound (26% yield). LCMS:511.20 (M+1)⁺; HPLC: 91.79% (@ 254 nm) (R_(t); 4.914); ¹H NMR (DMSO-d₆,400 MHz) δ 11.55 (s, 1H), 8.74 (d, 3H, J=4 Hz), 8.05 (s, 1H), 6.06 (s,1H), 5.89 (s, 1H), 5.15-5.08 (m, 1H), 4.36 (d, 2H, J=4.8 Hz), 2.59 (s,2H), 2.23 (s, 3H), 2.11 (s, 3H), 1.54 (s, 6H), 1.49 (s, 6H), 1.48 (d,6H, J=7.6 Hz).

Synthesis of Compound B-13:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-3-methyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: ethyl6-hydroxy-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A stirred solution of 5-methyl-1H-pyrazol-3-amine (5 g, 51.48 mmol) inacetic acid (33 mL) and water (100 mL) was cooled to 0° C. and diethyloxaloacetate sodium salt (10.81 g, 51.48 mmol) was added to it.Resulting solution was heated at 100° C. for overnight. After completionof reaction the solid was filtered and dried to obtain the desiredintermediate (2.5 g, 21.9%).

Step 2: Synthesis of ethyl6-bromo-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

6-Hydroxy-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (2.5 g, 11.3mmol) was suspended in acetonitrile (25 mL) and POBr₃ (6.5 g, 22.6 mmol)was added to it. The reaction mixture was refluxed for 6 h. Oncompletion of reaction, acetonitrile was removed under reduced pressureand residue neutralized with saturated NaHCO₃. Extraction was carriedout using EtOAc and the combined organic layers were washed with water,brine and dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography to provide the target compound (2.5 g, 77.8%).

Step 3: Synthesis of ethyl6-bromo-1-isopropyl-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1 g,3.52 mmol) was suspended in acetonitrile (10 mL) and K₂CO₃ (0.728 g,5.28 mmol) and 2-bromopropane (0.866 g, 7.04 mmol) was added to it. Thereaction mixture was refluxed for overnight. On completion of reaction,acetonitrile was removed under reduced pressure and water added to it.Extraction was carried out using ethyl acetate and the combined organiclayers were washed with water, brine and dried over anhydrous Na₂SO₄.Solvent was removed under reduced pressure and residue was purified bysilica gel column chromatography to obtain the desired intermediate(0.61 g, 53.5%). Structure was confirmed by NOE.

Step 4: Synthesis of6-bromo-1-isopropyl-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid

To a solution of ethyl6-bromo-1-isopropyl-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.6 g, 1.84 mmol) in EtOH (10 mL), aqueous NaOH (0.11 g, 2.76 mmol, in5 mL water) was added and reaction mixture stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and it was acidified using 10% citric acid solution. Extractionwas carried out using 5% MeOH/DCM. The combined organic layers werewashed with water & dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford target compound (0.35 g,63.8%), which was not purified further.

Step 5: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-1-isopropyl-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylicacid (0.3 g, 1.01 mmol) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.306 g, 2.01 mmol) inDMSO (4 mL) and was stirred at room temperature for 15 min. Then PyBOP(0.78 g, 1.51 mmol) was added to it and stirring was continued for 12 h.After completion of the reaction, reaction mass was poured into ice toobtain solid, which was filtered and washed with acetonitrile followedby ether to provide6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.12 g, 27.5%).

Step 6: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-3-methyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-3-methyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.1 g, 0.231 mmol), boronic ester (0.073 g, 0.277 mmol) and Pd(PPh₃)₄(0.013 g, 0.011 mmol) in 1,4-dioxane (1 mL) was purged with argon for 10min. Then, 2M Na₂CO₃ solution (0.08 g, 0.833 mmol) was added to it andagain argon was purged through it for 10 min. The reaction mixture wasstirred at 100° C. for 1 h. After completion of the reaction, water wasadded to it and extraction was carried out using 10% MeOH/DCM. Thecombined organic layers were washed with water, dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredfinal target (12% yield). LCMS: 491.35 (M+1)⁺; HPLC: 99.28% (@ 254 nm),(R_(t); 5.042; ¹H NMR (DMSO-d₆, 400 MHz) δ 11.46 (bs, 1H), 8.61 (t, 1H,J=4.4 Hz), 7.34 (s, 1H), 6.73 (s, 1H), 5.87 (s, 1H), 5.16-5.10 (m, 1H),4.36 (d, 2H, J=5.2 Hz), 2.38 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.45(d, 6H, J=6.8 Hz), 1.26 (s, 6H), 1.23 (s, 2H), 1.17 (s, 6H).

Synthesis of Compound B-14:1-cyclobutyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-bromo-1-cyclobutyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1 equiv.) wassuspended in acetonitrile and K₂CO₃ (1.5 equiv.) and bromocyclobutane (2equiv.) was added to it. The reaction mixture was refluxed for 8 h. Oncompletion, acetonitrile was removed under reduced pressure and waterwas added to it. Extraction was carried out using ethyl acetate; thecombined organic layers were washed with water, brine and dried overanhydrous Na₂SO₄. Solvent was removed under reduced pressure and residuewas purified by silica gel column chromatography to obtain the desiredintermediate.

Step 2: Synthesis of6-bromo-1-cyclobutyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (5 equivalent) was added to a solution of ethyl6-bromo-1-cyclobutyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1equivalent) in EtOH (10 times by volume) and stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and it was acidified using 10% citric acid solution. Extractionwas carried out using EtOAc; the combined organic layers were washedwith water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure. The residue was, then, dissolved inDMSO (10 times by volume) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equivalents) was addedto it. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (1.5 equivalents) was added to it and stirring wascontinued for 5 to 16 h. After completion of the reaction, water wasadded to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water; brine; dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography followed by ether washto provide the desired intermediate.

Step 3: Synthesis of1-cyclobutyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-1-cyclobutyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) solution was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound (80% yield). LCMS: 489.30 (M+1)⁺; HPLC: 95.97% (@ 254 nm)(R_(t); 5.425); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 8.89 (t, 1H),8.31 (s, 1H), 7.79 (s, 1H), 6.81 (s, 1H), 5.89 (s, 1H), 5.52-5.44 (m,1H), 4.37 (d, 2H, J=4.8 Hz), 2.73-2.63 (m, 2H), 2.43 (m, 4H), 2.22 (s,3H), 2.12 (s, 3H), 1.94-1.88 (m, 2H), 1.24 (d, 6H, J=6 Hz), 1.15 (s,6H).

Synthesis of Compound B-15:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(oxetan-3-yl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-bromo-1-(oxetan-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1 equiv.) wassuspended in acetonitrile and K₂CO₃ (1.5 equiv.) and 3-bromooxetane (2equiv.) was added to it. The reaction mixture was refluxed for 8 h. Oncompletion, acetonitrile was removed under reduced pressure and waterwas added to it. Extraction was carried out using ethyl acetate; thecombined organic layers were washed with water, brine and dried overanhydrous Na₂SO₄. Solvent was removed under reduced pressure and residuewas purified by silica gel column chromatography to obtain the desiredintermediate.

Step 2: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (5 equivalent) was added to a solution of ethyl6-bromo-1-(oxetan-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1equivalent) in EtOH (10 times by volume) and stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and it was acidified using 10% citric acid solution. Extractionwas carried out using EtOAc; the combined organic layers were washedwith water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure. The residue was, then, dissolved inDMSO (10 times by volume) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equivalents) was addedto it. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (1.5 equivalents) was added to it and stirring wascontinued for 5 to 16 h. After completion of the reaction, water wasadded to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water; brine; dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography followed by ether washto provide the desired intermediate.

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(oxetan-3-yl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) solution was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound (44% yield). LCMS: 491.50 (M+1)⁺; HPLC: 98.99% (@ 254 nm)(R_(t); 4.678); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.54 (bs, 1H), 8.93 (t,1H), 8.43 (s, 1H), 7.83 (s, 1H), 6.84 (s, 1H), 6.21-6.14 (m, 1H), 5.89(s, 1H), 5.08-5.02 (m, 4H), 4.38 (d, 2H, J=4.4 Hz), 2.42 (s, 2H), 2.23(s, 3H), 2.12 (s, 3H), 1.24 (s, 6H), 1.14 (s, 6H).

Synthesis of Compound B-16:1-cyclohexyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-bromo-1-cyclohexyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1 equiv.) wassuspended in acetonitrile and K₂CO₃ (1.5 equiv.) and bromocyclohexane (2equiv.) was added to it. The reaction mixture was refluxed for 8 h. Oncompletion, acetonitrile was removed under reduced pressure and waterwas added to it. Extraction was carried out using ethyl acetate; thecombined organic layers were washed with water, brine and dried overanhydrous Na₂SO₄. Solvent was removed under reduced pressure and residuewas purified by silica gel column chromatography to obtain the desiredintermediate.

Step 2: Synthesis of6-bromo-1-cyclohexyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (5 equivalent) was added to a solution of ethyl6-bromo-1-cyclohexyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (1equivalent) in EtOH (10 times by volume) and stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and it was acidified using 10% citric acid solution. Extractionwas carried out using EtOAc; the combined organic layers were washedwith water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure. The residue was, then, dissolved inDMSO (10 times by volume) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equivalents) was addedto it. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (1.5 equivalents) was added to it and stirring wascontinued for 5 to 16 h. After completion of the reaction, water wasadded to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water; brine; dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography followed by ether washto provide the desired intermediate.

Step 3: Synthesis of1-cyclohexyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-1-cyclohexyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) solution was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound (7% yield). LCMS: 517.35 (M+1)⁺; HPLC: 99.68% (@ 254 nm),(R_(t); 5.761); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.54 (bs, 1H), 8.86 (t,1H), 8.25 (s, 1H), 7.78 (s, 1H), 6.78 (s, 1H), 5.89 (s, 1H), 4.82-4.76(m, 1H), 4.38 (d, 2H, J=4.8 Hz), 2.22 (s, 3H), 2.12 (s, 3H), 2.01-1.85(m, 8H), 1.53-1.44 (m, 2H), 1.27 (bs, 6H), 1.23 (bs, 2H), 1.17 (bs, 6H).

Synthesis of Compound B-17:6-acetyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-ethoxyvinyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.1 g, 0.24 mmol) and tributyl(1-ethoxyvinyl)stannane (0.09 mL, 0.26mmol) in 1,4-dioxane (2 mL) was purged with argon for 10 min. Then,Pd(PPh₃)₄ (0.014 g, 0.012 mmol) was added to it and again argon waspurged through it for 10 min. The reaction mixture was refluxed for 4 h.After completion of the reaction, solvent was removed under reducedpressure to afford crude material which was as such used for next step(0.12 g).

Step 2: Synthesis of6-acetyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

35% HCl (4 mL) was added to theN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-ethoxyvinyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.12 g, 0.293 mmol) at 0° C. and stirred it at room temperature for 1h. After completion of reaction, saturated NaHCO₃ solution was added toit and extraction was carried out by using 5% MeOH/DCM, the combinedorganic layers were washed with water, brine and dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography to provide the desiredcompound (90% yield). LCMS: 382.10 (M+1)⁺; HPLC: 99.55% (@ 254 nm)(R_(t); 6.316); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 9.09 (bs,1H), 8.48 (s, 1H), 8.14 (s, 1H), 5.89 (s, 1H), 5.35-5.29 (m, 1H), 4.36(d, 2H, J=4.0 Hz), 2.75 (s, 3H), 2.21 (s, 3H), 2.12 (s, 3H), 1.56 (d,6H, J=6.8 Hz).

Synthesis of Compound B-18:N4-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-N6-methyl-1H-pyrazolo[3,4-b]pyridine-4,6-dicarboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-vinyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 g, 2.39 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (0.442g, 2.87 mmol) and Pd(PPh₃)₄ (0.276 g, 0.238 mmol) in 1,4-dioxane (10 mL)was purged with argon for 10 min. Then, solution of Na₂CO₃ (0.91 g, 8.58mmol) in water (4.3 mL) was added to it and again argon was purgedthrough it for 10 min. The reaction mixture was stirred at 100° C. for 1h. After completion of the reaction, water was added to it andextraction was carried out using 20% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give desired product (0.8 g,91.6%).

Step 2: Synthesis of6-(1,2-dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-vinyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.8 g, 2.19 mmol) in DCM (10 mL), N-Methylmorpholine-N-oxide (0.769 g,6.57 mmol) was added at 0° C. Then OsO₄ (2.5% in t-BuOH) (0.139 g, 5.56mL, 0.547 mmol) was added to it. Resulting solution was stirred at roomtemperature for 1 h. After completion of reaction, water was added to itand extraction was carried out using 20% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give the desired product (0.66 g,75.4%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-(1,2-dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.66 g, 1.65 mmol) in 50% THF/Water (12 mL), NaIO₄ (1.06 g, 4.95 mmol)was added at 0° C. and stirred at room temperature for 1 h. Aftercompletion of reaction, solid was filtered and washed with water.Azeotrope it with toluene to obtainN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.515 g, 84.8%).

Step 4: Synthesis of4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-6-carboxylicacid

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.515 g, 1.40 mmol) in DMF (3 mL), Oxone (0.861 g, 1.40 mmol) was addedand stirred it at room temperature for 3 h. On completion of reaction,water was added to it and extraction was carried out using 20% MeOH/DCM.The combined organic layers were washed with water, dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford crudematerial which was as such used for next step.

Step 5: Synthesis ofN4-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-N6-methyl-1H-pyrazolo[3,4-b]pyridine-4,6-dicarboxamide

4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-6-carboxylicacid (1 equiv.) was dissolved in DMSO and methylamine (2 equiv.) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (1.5 equiv.) was added to it and stirring was continuedovernight. After completion of the reaction, water was added to it andextraction was carried out using 20% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography/prep. HPLC to give the desiredcompound (8% yield). LCMS: 397.15 (M+1)⁺; HPLC: 93.04% (@ 254 nm)(R_(t); 5.597); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 9.04 (t, 1H,J=4.4 Hz), 8.95 (d, 1H, J=4.8 Hz), 8.43 (s, 1H), 8.17 (s, 1H), 5.88 (s,1H), 5.49-5.43 (m, 1H), 4.36 (d, 2H, J=4.8 Hz), 2.88 (d, 3H, J=4.4 Hz),2.21 (s, 3H), 2.12 (s, 3H), 1.52 (d, 6H, J=6.8 Hz).

Synthesis of Compound B-19:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(1-phenylethyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Aqueous NaOH (1.11 g, 27.88 mmol) was added to a solution of ethyl6-bromo-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (5 g, 18.6 mmol) inEtOH (50 mL) and stirred at 60° C. for 1 h. After completion of thereaction, ethanol was removed under reduced pressure and it wasacidified using 10% citric acid solution. Solid obtained was filteredand dried under reduced pressure. The crude acid (4.4 g, 18.3 mmol) wasthen dissolved in DMSO (20 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (5.55 g, 36.51 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (14.24 g, 27.38 mmol) was added to it and stirring wascontinued overnight. After completion of the reaction, reaction mass waspoured into ice to obtain solid, which was filtered and washed withacetonitrile followed by ether to provide the desired intermediate (4 g,58.47%).

Step 2: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(1-phenylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

6-Bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.25 g, 0.66 mmol) was dissolved in DMF (3 mL) and K₂CO₃ (0.17 g, 0.79mmol) and (1-bromoethyl)benzene (0.148 g, 0.799 mmol) was added to it.The reaction mixture was refluxed for overnight. On completion, waterwas added to it. Extraction was carried out using DCM; the combinedorganic layers were washed with water, brine and dried over anhydrousNa₂SO₄. Solvent was removed under reduced pressure and residue waspurified by silica gel column chromatography to obtain the desiredintermediate (0.26 g, 92.8%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(1-phenylethyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(1-phenylethyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.26 g, 0.54 mmol),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(0.172 g, 0.649 mmol.) and Pd(PPh₃)₄ (0.062 g, 0.054 mmol) in1,4-dioxane (3 mL) was purged with argon for 10 min. Then, 2 M Na₂CO₃(0.206 g, 1.94 mmol.) in water was added to it and again argon waspurged through it for 10 min. The reaction mixture was stirred at 100°C. for 2 h. After completion of the reaction, water was added to it andextraction was carried out using 20% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by prep. HPLC to give the desired compound (12% yield). LCMS539.20 (M+1)⁺; HPLC: 97.09% (@ 254 nm) (R_(t); 5.747); ¹H NMR (DMSO-d₆,400 MHz) δ 11.54 (bs, 1H), 8.84 (t, 1H, J=5.2 Hz), 8.73-8.64 (m, 2H),8.36 (s, 1H), 7.86 (s, 1H), 7.34-7.21 (m, 5H), 6.84 (s, 1H), 6.30 (q,1H, J=6.8 Hz), 5.89 (s, 1H), 4.38 (d, 2H, J=5.2 Hz), 2.88-2.77 (m, 2H),2.23 (s, 3H) 2.12 (s, 3H), 1.95 (d, 3H, J=7.2 Hz), 1.56 (s, 6H), 1.46(s, 6H).

Synthesis of Compound B-20:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was dissolved in DMSO, to which Et₃N (3 equiv.) and1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (2 equiv.) wereadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (1.5 equiv.) was added to it and stirring was continuedovernight. After completion of the reaction, water was added to it.Extraction was carried out using DCM; the combined organic layers werewashed with water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure to afford crude material which wastaken up in purified by column chromatography. The Boc protecting groupwas removed using standard conditions to give the desired compound (83%yield). LCMS: 532.35 (M+1)⁺; HPLC: 99.04% (@ 254 nm) (R_(t); 5.006); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.54 (bs, 1H), 8.83 (t, 1H), 8.47 (bs, 1H),8.30 (s, 1H), 8.26 (bs, 1H), 7.86 (s, 1H), 6.92 (bs, 1H), 5.89 (s, 1H),5.21-5.18 (m, 1H), 4.38 (d, 2H, J=4 Hz), 4.22 (bs, 2H), 3.75 (m, 2H),3.32 (m, 3H), 2.96 (m, 2H), 2.80 (bs, 1H), 2.67 (bs, 1H), 2.20 (s, 3H),2.12 (s, 3H), 1.84-1.73 (m, 4H), 1.51 (d, 6H, J=6.8 Hz).

Synthesis of Compound B-21:1-isopropyl-N-((4-(methoxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of 5-methoxypent-3-yn-2-one

A stirred solution of 3-methoxyprop-1-yne (3 g, 42.8 mmol) in THF (60mL) was cooled to 0° C. and n-BuLi (32 mL, 51.2 mmol, 1.6 M in hexanes)was added to it. After addition, reaction was gradually warmed to roomtemperature over a period of 2 h. The reaction was again cooled to −78°C. and BF₃OEt₂ (7.28 g, 51.3 mmol) was added to it. After 5 min aceticanhydride (5.72 g, 56.1 mmol) was added. The reaction was graduallywarmed to room temperature over a period of 2 h. After this time, 1NNaOH was added to reaction mixture until solution become neutral. Thebiphasic solution was extracted with diethyl ether. Combined organiclayers were dried over sodium sulfate and concentrated to obtain crudematerial which was purified by column over silica gel affording thedesired compound (1.2 g, 24%).

Step 2: Synthesis of4-(methoxymethyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile

To a solution of 5-methoxypent-3-yn-2-one (1.2 g, 10.7 mmol) in 90%ethanol (22 mL), cyanoacetamide (1.08 g, 12.85 mmol) and piperidineacetate {prepared by the addition of piperidine to a solution of aceticacid (0.5 mL) in water (1 mL) till pH 8} were added and reaction heatedat 90° C. for 16 h. On completion, ethanol was evaporated and water wasadded to residue. Solid obtained was filtered and washed with water.Then this solid was stirred with acetonitrile for 10 min and filteredagain to obtain the desired intermediate (0.165 g, 8.68%).

Step 3: Synthesis of3-(aminomethyl)-4-(methoxymethyl)-6-methylpyridin-2(1H)-one

To a solution of4-(methoxymethyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile(0.165 g, 0.927 mmol) in methanol (10 mL), Raney Ni and ammonia (1 mL)were added and reaction stirred under hydrogen balloon pressure for 3 h.On completion, reaction mass was filtered through celite bed andfiltrate concentrated to obtain the desired compound (0.13 g, 80%).

Step 4: Synthesis of6-bromo-1-isopropyl-N-((4-(methoxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a solution of6-bromo-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid (0.155g, 0.549 mmol) in DMSO (2 mL), PyBOP (0.429 g, 0.823 mmol) was added andreaction stirred at room temperature for 15 min. Then3-(aminomethyl)-4-(methoxymethyl)-6-methylpyridin-2(1H)-one (0.1 g,0.549 mmol) was added and reaction stirred overnight. On completion,water was added and solid that precipitates out was filtered and washedwith water to obtain the desired intermediate (0.06 g, 24.5%).

Step 5: Synthesis of1-isopropyl-N-((4-(methoxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-bromo-1-isopropyl-N-((4-(methoxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.06 g, 0.133 mmol) and(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)boronic acid (0.043g, 0.160 mmol) in dioxane/water mixture (1.5 mL+0.5 mL), Na₂CO₃ (0.5 g,0.5 mmol) was added and solution purged with argon for 15 min. ThenPd(PPh₃)₄ (0.008 g, 0.007 mmol) was added and argon was purged again for10 min. Reaction mass was heated at 100° C. for 2 h. On completion,reaction mixture was diluted with water and extracted with 10% MeOH/DCM.Combined organic layers were dried over Na₂SO₄ and solvent removed underreduced pressure to obtain crude material which was purified by columnchromatography over silica gel to afford the desired product (44%yield). LCMS: 507.25 (M+1)⁺; HPLC: 99.64% (@ 254 nm) (R_(t); 5.159); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.68 (bs, 1H), 8.91 (t, 1H), 8.26 (s, 1H),7.77 (s, 1H), 6.80 (s, 1H), 6.09 (s, 1H), 5.24-5.18 (m, 1H), 4.51 (s,2H), 4.35 (d, 2H, J=4.4 Hz), 3.32 (3H merged in DMSO peak), 2.43 (s,2H), 2.17 (s, 3H), 1.49 (d, 6H, J=6 Hz), 1.24 (s, 6H), 1.14 (s, 6H).

Synthesis of Compound B-22:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-vinyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 g, 2.39 mmol), boronic ester (0.442 g, 2.87 mmol) and Pd(PPh₃)₄(0.276 g, 0.238 mmol) in 1,4-dioxane (10 mL) was purged with argon for10 min. Then, solution of Na₂CO₃ (0.91 g, 8.58 mmol) in water (4.3 mL)was added to it and again argon was purged through it for 10 min. Thereaction mixture was stirred at 100° C. for 1 h. After completion of thereaction, water was added to it and extraction was carried out using 20%MeOH/DCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound (0.8 g, 91.6%).

Step 2: Synthesis of6-(1,2-dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-vinyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.8 g, 2.19 mmol) in DCM (10 mL), N-Methylmorpholine-N-oxide (0.769 g,6.57 mmol) was added at 0° C. Then 2.5% OsO₄ in t-BuOH (0.139 g, 5.56mL, 0.547 mmol) was added to it. Resulting solution was stirred at roomtemperature for 1 h. After completion of reaction, water was added to itand extraction was carried out using 20% MeOH/DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give the desired compound (0.66 g,75.4%).

Step 3: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-(1,2-dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide0.66 g, 1.65 mmol) in 50% THF/Water (12 mL), NaIO₄ (1.06 g, 4.95 mmol)was added at 0° C. and stirred at room temperature for 1 h. Aftercompletion of reaction, solid was filtered and washed with water.Azeotrope with toluene to obtain the desired compound (0.515 g, 84.8%).LCMS: 368.05 (M+1)⁺; HPLC: 99.53% (@ 254 nm), (R_(t); 5.901); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.54 (s, 1H), 10.09 (s, 1H), 9.10 (bs, 1H), 8.53(s, 1H), 8.11 (s, 1H), 5.89 (s, 1H), 5.37-5.31 (m, 1H), 4.37 (d, 2H,J=4.4 Hz), 2.21 (s, 3H), 2.12 (s, 3H), 1.55 (d, 6H, J=6.4 Hz).

Synthesis of Compound B-23:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethylpiperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(1.2 equiv.) and Pd(PPh₃)₄ (0.1 equiv.) in 1,4-dioxane was purged withargon for 10 min. Then, 2 M Na₂CO₃ (3.6 equiv.) in water was added to itand again argon was purged through it for 10 min. The reaction mixturewas stirred at 100° C. for 1 h. After completion of the reaction, waterwas added to it and extraction was carried out using EtOAc. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give the desiredintermediate.

Step 2:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethylpiperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.07 g, 0.147 mmol) in MeOH (2 mL) was added 10% Pd/C in catalyticamount and stirred it at room temperature under hydrogen pressure(bladder pressure) for 3 hr. On completion of reaction, filter itthrough celite bed, then filtrate was concentrated under reducepressure. The crude material was purified by column chromatography togive the desired compound (71% yield). LCMS: 479.20 (M+1)⁺; HPLC: 99.04%(@ 254 nm), (R_(t); 5.075); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H),8.70 (t, 1H), 8.27 (s, 1H), 7.70 (bs, 1H), 7.53 (s, 1H), 5.89 (s, 1H),5.22-5.15 (m, 1H), 4.38 (d, 2H, J=5.2 Hz), 3.57 (m, 1H), 2.22 (s, 3H),2.12 (s, 3H), 2.00-1.90 (m, 2H), 1.90-1.80 (m, 2H), 1.50 (s, 6H), 1.49(s, 9H), 1.40 (s, 3H).

Synthesis of Compound B-24:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(piperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl6-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A solution of ethyl1-isopropyl-6-(((trifluoromethyl)sulfonyl)oxy)-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.45 g, 1.181 mmol), boronic acid (0.4 g, 1.299 mmol) and Pd(PPh₃)₄(0.137 g, 0.118 mmol) in 1,4-dioxane (6 mL) was purged with argon for 10min. Then, 2 M Na₂CO₃ (0.45 g, 4.252 mmol) was added to it and againargon was purged through it for 10 min. The reaction mixture was stirredat 100° C. for 1 h. After completion of the reaction, water was added toit and extraction was carried out using EtOAc. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give the desired compound (0.45 g,92.0%).

Step 2: Synthesis of ethyl6-(1-(tert-butoxycarbonyl)piperidin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

To a stirred solution of ethyl6-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.45 g, 1.086 mmol) in EtOH (10 mL), 10% Pd/C in catalytic amount wasadded and stirred it at room temperature under hydrogen pressure(bladder pressure) for 3 hr. On completion of reaction, reaction masswas filtered through celite bed, then filtrate was concentrated underreduce pressure to give the desired compound (0.34 g, 75%).

Step 3: Synthesis of tert-butyl4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridin-6-yl)piperidine-1-carboxylate

Aqueous NaOH (0.065 g, 1.634 mmol) was added to a solution of ethyl6-(1-(tert-butoxycarbonyl)piperidin-4-yl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.34 g, 0.817 mmol) in EtOH (4 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand it was acidified using 10% citric acid solution. Extraction wascarried out using ethyl acetate; the combined organic layers were washedwith water, brine and dried over anhydrous Na₂SO₄; filtered andconcentrated under reduced pressure. The crude acid (0.3 g, 0.773 mmol)was then dissolved in DMSO (3 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.235 g, 1.546 mmol) wasadded to it. The reaction mixture was stirred at room temperature for 15min before PYBOP (0.603 g, 1.159 mmol) was added to it and stirring wascontinued overnight. After completion of the reaction, reaction mass waspoured into ice to obtain solid, which was filtered and washed withacetonitrile followed by ether to provide the desired intermediate (0.2g, 50.0%).

Step 4: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(piperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Tert-butyl 4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridin-6-yl)piperidine-1-carboxylate(0.2 g, 0.383 mmol) was taken in DCM (3 mL), to it TFA (0.6 mL) wasadded at 0° C. and stirred at room temperature for 3 h. On completion ofreaction, DCM was removed under reduced pressure, saturated NaHCO₃solution was added to it and extraction was carried out using DCM. Thecombined organic layers were washed with water, dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give the desiredcompound (60% yield). LCMS: 423.25 (M+1)⁺; HPLC: 99.121% (@ 254 nm)(R_(t); 4.628); ¹H NMR (DMSO-d₆, 400 MHz) δ 8.75 (t, J=4.8 Hz, 1H); 8.26(s, 1H); 7.49 (s, 1H); 5.89 (s, 1H); 5.22-5.16 (m, 1H); 4.36 (d, J=4.8Hz, 2H); 3.04 (d, J=12.0 Hz, 2H); 2.84-2.92 (m, 1H); 2.60 (t, J=, 8.4Hz, 2H); 2.20 (s, 3H); 2.10 (s, 3H); 1.65-1.81 (m, 4H); 1.49 (d, J=6.8Hz, 6H).

Synthesis of Compound B-25:N-((4-ethyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of 4-ethyl-2-hydroxy-6-methylnicotinonitrile

To a solution of hex-3-yn-2-one (1.5 g, 15.6 mmol) in 90% ethanol (30mL), cyanoacetamide (1.58 g, 18.72 mmol) and piperidine acetate{prepared by the addition of piperidine to a solution of acetic acid(0.258 mL) in water (0.65 mL) till pH 8} were added and reaction heatedat 90° C. for 12 h. On completion, ethanol was evaporated and water wasadded to residue. Solid obtained was filtered and washed with water.Then this solid was stirred with acetonitrile for 10 min and filteredagain to obtain the desired intermediate (1.5 g, 60%).

Step 2: Synthesis of 3-(aminomethyl)-4-ethyl-6-methylpyridin-2-ol

To a solution of 4-ethyl-2-hydroxy-6-methylnicotinonitrile (0.5 g, 3.08mmol) in methanol (10 mL), Raney Ni and ammonia (1 mL) were added andreaction stirred under hydrogen balloon pressure for 3 h. On completion,reaction mass was filtered through celite bed and filtrate concentratedto obtain the desired product (0.45 g, 87.9%).

Step 3: Synthesis of6-bromo-N-((4-ethyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in DMSO (3 mL for 1 mmol), PyBOP (1.5 equiv.) was added andreaction stirred at room temperature for 15 min. Then3-(aminomethyl)-4-ethyl-6-methylpyridin-2(1H)-one (2 equiv.) was addedand reaction stirred overnight. On completion, water was added and solidthat precipitates out was filtered and washed with water. Then thissolid was stirred with acetonitrile for 10 min and filtered again toobtain the desired intermediate.

Step 4: Synthesis ofN-((4-ethyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution of6-bromo-N-((4-ethyl-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.37 g, 0.85 mmol) and2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(1.2 equiv.) in dioxane/water mixture (3 mL+1 mL for 1 mmol), Na₂CO₃(3.6 equiv.) was added and solution purged with argon for 15 min. ThenPd(PPh₃)₄ (0.05 equiv.) was added and argon was purged again for 10 min.Reaction mass was heated at 100° C. for 2 h. On completion, reactionmixture was diluted with water and extracted with 10% MeOH/DCM. Combinedorganic layers were dried over Na₂SO₄ and solvent removed under reducedpressure to afford crude material which was purified by columnchromatography over silica gel to afford the desired compound. (38%yield). LCMS: 491.25 (M+1)⁺; HPLC: 95.01% (@ 254 nm) (R_(t); 5.463); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 8.87 (s, 1H), 8.26 (s, 1H), 7.79(s, 1H), 6.80 (s, 1H), 5.91 (s, 1H), 5.22-5.19 (m, 1H), 4.39 (d, 2H,J=4.0 Hz), 2.42-2.40 (m, 4H), 2.24 (s, 3H), 1.49 (d, 6H, J=6 Hz). 1.24(s, 6H), 1.14 (s, 6H), 1.11 (m, 3H).

Synthesis of Compound B-26:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-ethyl-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of ethyl3-ethyl-6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

A stirred solution of 5-ethyl-1H-pyrazol-3-amine (1 g, 9.00 mmol) inacetic acid (6.6 mL) and water (20 mL) was cooled to 0° C. and diethyloxaloacetate sodium salt (1.88 g, 9.00 mmol) was added to it. Resultingsolution was heated at 100° C. for overnight. After completion of thereaction, the solid was filtered and dried to obtain the desiredintermediate (0.52 g, 24.6%).

Step 2: Synthesis of ethyl6-bromo-3-ethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 3-ethyl-6-hydroxy-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (0.52g, 2.21 mmol) was suspended in acetonitrile (6 mL) and POBr₃ (1.27 g,4.42 mmol) was added to it. The reaction mixture was refluxed for 6 h.On completion of reaction, acetonitrile was removed under reducedpressure and residue neutralized with saturated NaHCO₃. Extraction wascarried out using EtOAc; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography to provide the desired intermediate (0.45 g, 68.5%).

Step 3: Synthesis of ethyl6-bromo-3-ethyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate

Ethyl 6-bromo-3-ethyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate (0.45 g,1.51 mmol) was suspended in acetonitrile (5 mL) and K₂CO₃ (0.312 g, 2.26mmol) and 2-bromopropane (0.372 g, 3.02 mmol) was added to it. Thereaction mixture was refluxed for overnight. On completion of reaction,acetonitrile was removed under reduced pressure and water was added toit. Extraction was carried out using ethyl acetate; the combined organiclayers were washed with water, brine and dried over anhydrous Na₂SO₄.Solvent was removed under reduced pressure and residue was purified bysilica gel column chromatography to give the desired intermediate (0.4g, 87.7%).

Step 4: Synthesis of6-bromo-3-ethyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid

To a solution of ethyl6-bromo-3-ethyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylate(0.4 g, 1.176 mmol) in EtOH (7 mL), aqueous NaOH (0.062 g, 1.529 mmol in1 mL water) was added and reaction mixture stirred at 60° C. for 1 h.After completion of the reaction, ethanol was removed under reducedpressure and it was acidified using 2N HCl and 10% citric acid solution.Solid obtained was filtered and azeotrope it with toluene to give thedesired compound (0.3 g, 81.7%).

Step 5: Synthesis of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-ethyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-3-ethyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxylic acid(0.3 g, 0.901 mmol) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one(0.292 g, 1.92 mmol) in DMSO (4 mL) and was stirred at room temperaturefor 15 min. Then PyBOP (0.75 g, 1.44 mmol) was added to it and stirringwas continued for 12 hr. After completion of the reaction, reaction masswas poured into ice to obtain solid, which was filtered and washed withacetonitrile followed by ether to provide the desired intermediate (0.35g, 81.6%).

Step 6: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-ethyl-1-isopropyl-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-ethyl-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.15 g, 0.336 mmol),2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(0.106 g, 0.403 mmol) and Pd(PPh₃)₄ (0.020 g, 0.017 mmol) in 1,4-dioxane(3 mL) was purged with argon for 10 min. Then, 2M Na₂CO₃ solution (0.128g, 1.21 mmol) was added to it and again argon was purged through it for10 min. The reaction mixture was stirred at 100° C. for 1 h. Aftercompletion of the reaction, water was added to it and extraction wascarried out using 10% MeOH/DCM. The combined organic layers were washedwith water, dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to afford crude material which was purified by columnchromatography to give the desired compound (59% yield). LCMS: 505.30(M+1)⁺; HPLC: 99.84% (@ 254 nm) (R_(t); 5.386); ¹H NMR (DMSO-d₆, 400MHz) δ 11.49 (s, 1H), 8.68 (t, 1H), 7.32 (s, 1H), 6.72 (s, 1H), 5.88 (s,1H), 5.17-5.10 (m, 1H), 4.33 (d, 2H, J=4.4 Hz), 2.84-2.79 (q, 2H, J=7.6Hz), 2.39 (s, 2H), 2.25 (s, 3H), 2.11 (s, 3H), 1.46 (d, 6H, J=6.8 Hz),1.22 (s, 6H), 1.13 (s, 6H), 1.06 (t, 3H, J=7.6 Hz).

Synthesis of Compound B-27:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(methylsulfonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.2 g, 0.476 mmol) in pyridine (1.5 mL), mesyl chloride (0.081 g, 0.714mmol) was added at 0° C. and stirred it at room temperature for 1 h.After completion of reaction, solvent was removed under reduced pressureand water was added to it. Extraction was carried out using DCM; thecombined organic layers were washed with water, brine and dried overanhydrous Na₂SO₄; filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound. LCMS: 499.20 (M+1)⁺; HPLC: 98.22% (@ 254 nm)(R_(t); 6.278); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 8.85 (t, 1H),8.31 (s, 1H), 7.86 (s, 1H), 6.94 (s, 1H), 5.89 (s, 1H), 5.22 (m, 1H),4.38 (d, 2H, J=4.8 Hz), 3.98 (m, 2H), 3.42 (t, 2H, J=5.6 Hz), 2.96 (s,3H), 2.84 (m, 2H), 2.21 (s, 3H), 2.13 (s, 3H), 1.50 (d, 6H, J=6.8 Hz).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(methylsulfonyl)piperidin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(methylsulfonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.08 g, 0.16 mmol) in EtOH (2 mL), 10% Pd/C in catalytic amount wasadded and stirred it at room temperature under hydrogen pressure(balloon pressure) for 3-4 hr. On completion of reaction, reaction masswas filtered through celite bed, then filtrate was concentrated underreduce pressure, finally the residue was washed with diethyl ether toprovide the desired compound (12% yield). LCMS: 501.15 (M+1)⁺; HPLC:94.05% (@ 254 nm) (R_(t); 6.087); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s,1H), 8.77 (t, 1H, J=4.8 Hz), 8.29 (s, 1H), 7.56 (s, 1H), 5.89 (s, 1H),5.21-5.17 (m, 1H), 4.37 (d, 2H, J=4.4 Hz), 3.69 (m, 2H), 3.01-2.96 (m,1H), 2.91 (s, 3H), 2.87-2.84 (m, 2H), 2.20 (s, 3H), 2.12 (s, 3H),2.08-1.99 (m, 2H), 1.93-1.83 (m, 2H), 1.49 (d, 6H, J=6.8 Hz).

Synthesis of Compound B-28:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was dissolved in DMSO, to it Et₃N (3 equiv.) andpiperidine-3-carboxylic acid (2 equiv.) was added to it. The reactionmixture was stirred at room temperature for 15 min before PYBOP (1.5equiv.) was added to it and stirring was continued overnight. Aftercompletion of the reaction, water was added to it. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to afford crude material which was purified bycolumn chromatography to give the desired compound (85% yield). LCMS:532.35 (M+1)⁺; HPLC: 99.84% (@ 254 nm) (R_(t); 5.059); ¹H NMR (DMSO-d₆,400 MHz) δ 11.55 (bs, 1H), 8.83 (t, 1H), 8.44 (bs, 2H), 8.30 (s, 1H),7.86 (s, 1H), 6.92 (bs, 1H), 5.89 (s, 1H), 5.21-5.18 (m, 1H), 4.38 (d,2H, J=4.4 Hz), 4.31 (bs, 1H), 4.23 (bs, 1H), 3.74 (m, 2H), 3.14 (m, 4H),2.97 (m, 1H), 2.82 (m, 1H), 2.70 (m, 1H), 2.20 (s, 3H), 2.12 (s, 3H),1.89 (m, 1H), 1.74 (m, 2H), 1.60 (m, 1H), 1.51 (d, 6H, J=6.4 Hz).

Synthesis of Compound B-29: of6-([1,4′-bipiperidin]-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis of tert-butyl4-(4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridin-6-yl)-5,6-dihydropyridin-1(2H)-yl)piperidine-1-carboxylate

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 g, 2.38 mmol) in methanol (15 mL) was added tert-butyl4-oxopiperidine-1-carboxylate (1.18 g, 5.95 mmol) and acetic acid (0.17g, 2.85 mmol) stirred it at room temperature for 2 days. Then NaBH₃CN(0.224 g, 3.57 mmol) was added to it resulting reaction mixture wasstirred at room temperature for 8 h. After completion of reaction,solvent was removed under reduced pressure and water was added to it.Extraction was carried out using ethyl acetate. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give the desired compound (0.5 g,34.5%).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Tert-butyl 4-(4-(4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridin-6-yl)-5,6-dihydropyridin-1(2H)-yl)piperidine-1-carboxylate(0.5 g, 0.829 mmol) was taken in DCM (6 mL), TFA (2 mL) was added andstirred at room temperature for 1 h. After completion of reaction,solvent was removed under reduced pressure, neutralized with saturatedNaHCO₃ solution. Extraction was carried out using DCM; the combinedorganic layers were washed with water, brine and dried over anhydrousNa₂SO₄; filtered and concentrated under reduced pressure, finally theresidue was washed with acetonitrile followed by diethyl ether toprovide the desired compound. LCMS: 504.35 (M+1)⁺; HPLC: 99.26% (@ 254nm) (R_(t); 4.223); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.56 (bs, 1H), 10.25(bs, 1H), 8.87 (t, 1H, J=4.8 Hz), 8.80 (bs, 1H), 8.55 (bs, 1H), 8.34 (s,1H), 7.89 (s, 1H), 6.93 (bs, 1H), 5.90 (s, 1H), 5.21 (m, 1H), 4.39 (d,2H, J=4.8 Hz), 4.08 (bs, 2H), 3.75 (bs, 2H), 3.30 (m, 3H), 3.16-3.12 (m,1H), 2.96 (m, 3H), 2.32 (m, 2H), 2.21 (s, 3H), 2.12 (s, 3H), 1.86 (m,2H), 1.52 (d, 6H, J=6.8 Hz).

Step 3: Synthesis of6-([1,4′-bipiperidin]-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.08 g, 0.159 mmol) in EtOH (2 mL), 10% Pd/C in catalytic amount wasadded and stirred it at room temperature under hydrogen pressure(balloon pressure) for 3-4 hr. On completion of reaction, reaction masswas filtered through celite bed, then filtrate was concentrated underreduce pressure, finally the residue was washed with diethyl ether toprovide the desired compound (62% yield). LCMS: 506.40 (M+1)⁺; HPLC:90.48% (@ 254 nm) (R_(t); 4.196); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (bs,1H), 9.79 (bs, 1H), 8.76 (t, 1H), 8.58 (bs, 1H), 8.27 (s, 1H), 7.47 (s,1H), 5.90 (s, 1H), 5.17 (m, 1H), 4.37 (d, 2H, J=4.4 Hz), 3.60 (bs, 2H),3.45 (bs, 2H), 3.16 (m, 4H), 2.95 (m, 4H), 2.21 (s, 3H), 2.12 (s, 3H),2.25-2.05 (m, 4H), 1.85 (m, 2H), 1.50 (d, 6H, J=6.4 Hz).

Synthesis of Compound B-30:6-(1-(azetidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was dissolved in DMSO, to it Et₃N (3 equiv.) andazetidine-3-carboxylic acid (2 equiv.) was added to it. The reactionmixture was stirred at room temperature for 15 min before PYBOP (1.5equiv.) was added to it and stirring was continued overnight. Aftercompletion of the reaction, water was added to it. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to afford crude material which was purified bycolumn chromatography to give the desired compound (81.9% yield). LCMS:504.25 (M+1)⁺; HPLC: 99.74% (@ 254 nm) (R_(t); 4.829); ¹H NMR (DMSO-d₆,400 MHz) δ 11.55 (bs, 1H), 8.84 (m, 2H), 8.64 (bs, 1H), 8.30 (s, 1H),7.86 (s, 1H), 6.93 (s, 1H), 5.89 (s, 1H), 5.22-5.17 (m, 1H), 4.38 (d,2H, J=4.8 Hz), 4.26 (bs, 1H), 4.20-4.10 (m, 4H), 4.05-3.97 (m, 1H), 3.75(t, 1H, J=5.2 Hz), 3.49 (m, 2H), 2.74 (m, 2H), 2.20 (s, 3H), 2.12 (s,3H), 1.51 (d, 6H, J=6.4 Hz).

Synthesis of Compound B-31:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylpyrrolidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(pyrrolidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was dissolved in DMSO, to it Et₃N (3 equiv.) andpyrrolidine-3-carboxylic acid (2 equiv.) was added to it. The reactionmixture was stirred at room temperature for 15 min before PYBOP (1.5equiv.) was added to it and stirring was continued overnight. Aftercompletion of the reaction, water was added to it. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to afford crude material which was purified bycolumn chromatography to give the desired compound.

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylpyrrolidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(pyrrolidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in methanol was added formalin (5 equiv.) stirred it at roomtemperature for 10 min. Then NaBH₃CN (1 equiv.) was added to itResulting reaction mixture was stirred at room temperature for 1 h.After completion of reaction, solvent was removed under reduced pressureand water was added to it. Extraction was carried out using 10%MeOH/DCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound (30% yield). LCMS: 532.35 (M+1)⁺; HPLC: 89.75%(@ 254 nm) (R_(t); 7.188); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H),8.84 (bs, 1H), 8.30 (s, 1H), 7.84 (d, 1H, J=10 Hz), 6.91 (bs, 1H), 5.89(s, 1H), 5.21 (m, 1H), 4.38 (d, 2H, J=4.4 Hz), 4.29 (bs, 1H), 4.21 (bs,1H), 3.70 (bs, 2H), 2.76 (m, 2H), 2.66 (bs, 1H), 2.43 (m, 1H), 2.32 (m,1H), 2.22 (s, 3H), 2.20 (s, 3H), 2.12 (s, 3H), 2.00-1.94 (m, 4H), 1.50(d, 6H, J=6.8 Hz).

Synthesis of Compound B-32:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylpiperidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) was dissolved in DMSO, to it Et₃N (3 equiv.) andpiperidine-3-carboxylic acid (2 equiv.) was added to it. The reactionmixture was stirred at room temperature for 15 min before PYBOP (1.5equiv.) was added to it and stirring was continued overnight. Aftercompletion of the reaction, water was added to it. Extraction wascarried out using DCM; the combined organic layers were washed withwater, brine and dried over anhydrous Na₂SO₄; filtered and concentratedunder reduced pressure to afford crude material which was purified bycolumn chromatography to give the desired compound.

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylpiperidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidine-3-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in methanol was added formalin (5 equiv.) stirred it at roomtemperature for 10 min. Then NaBH₃CN (1 equiv.) was added to itResulting reaction mixture was stirred at room temperature for 1 h.After completion of reaction, solvent was removed under reduced pressureand water was added to it. Extraction was carried out using 10%MeOH/DCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound (195 yield). LCMS: 546.40 (M+1)⁺; HPLC: 94.91%(@ 254 nm) (R_(t); 7.295); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (s, 1H),8.84 (bs, 1H), 8.30 (d, 1H, J=2 Hz), 7.84 (d, 1H, J=9.2 Hz), 6.90 (s,1H), 5.89 (s, 1H), 5.20 (m, 1H), 4.38 (d, 2H, J=4 Hz), 4.33 (bs, 1H),4.19 (bs, 1H), 3.72 (bs, 2H), 2.90-2.65 (m, 5H), 2.20 (s, 3H), 2.15 (s,3H), 2.12 (s, 3H), 1.91 (m, 1H), 1.80-1.70 (m, 3H), 1.61 (m, 1H), 1.50(d, 6H, J=6 Hz), 1.28 (m, 1H).

Synthesis of Compound B-33:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-(2-(methylsulfonyl)ethyl)piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.2 g, 0.397 mmol) in MeOH (2 mL), methyl vinyl sulfone (0.063 g, 0.596mmol) was added and stirred it at room temperature for 12 h. Aftercompletion of reaction, solvent was removed under reduced pressure andwater was added to it. Extraction was carried out using 10% MeOH/DCM;the combined organic layers were washed with water, brine and dried overanhydrous Na₂SO₄; filtered and concentrated under reduced pressure,finally the residue was washed with diethyl ether to provide the desiredcompound (41% yield). LCMS: 610.45 (M+1)⁺; HPLC: 97.26% (@ 254 nm)(R_(t); 6.037); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.54 (s, 1H), 8.82 (t, 1H),8.27 (s, 1H), 7.80 (s, 1H), 6.88 (s, 1H), 5.89 (s, 1H), 5.21-5.17 (m,1H), 4.37 (d, 2H, J=4.4 Hz), 3.32-3.25 (m, 4H), 3.02 (s, 3H), 2.95 (d,2H, J=10.40 Hz), 2.73 (d, 2H, J=4 Hz), 2.67 (bs, 4H), 2.32 (m, 2H), 2.20(s, 3H), 2.12 (s, 3H), 1.99-1.94 (m, 2H), 1.81-1.79 (m, 2H), 1.49 (d,6H, J=6.4 Hz), 1.45 (m, 1H).

Synthesis of Compound B-34:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-methylpiperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solutionN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(1 equiv.) in methanol was added formalin (5 equiv.) stirred it at roomtemperature for 10 min. Then NaBH₃CN (1 equiv.) was added to itResulting reaction mixture was stirred at room temperature for 1 h.After completion of reaction, solvent was removed under reduced pressureand water was added to it. Extraction was carried out using 10%MeOH/DCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive the desired compound (29% yield). LCMS: 546.30 (M+1)⁺; HPLC: 99.94%(@ 254 nm) (R_(t); 5.067); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (bs, 1H),8.83 (bs, 1H), 8.30 (s, 1H), 7.83 (d, 1H, J=13.20 Hz), 6.90 (bs, 1H),5.89 (s, 1H), 5.25-5.15 (m, 1H), 4.38 (d, 2H, J=3.6 Hz), 4.31 (bs, 1H),4.20 (bs, 1H), 3.71 (bs, 2H), 2.77 (bs, 3H), 2.65 (bs, 2H), 2.20 (s,3H), 2.14 (s, 3H), 2.12 (s, 3H), 1.91 (bs, 2H), 1.60 (bs, 4H), 1.50 (d,6H, J=6.4 Hz).

Synthesis of Compound B-35:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1′-(methylsulfonyl)-[1,4′-bipiperidin]-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

Step 1: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-(methylsulfonyl)piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.2 g, 0.397 mmol) in DCM (5 mL), Et₃N (0.06 g, 0.596 mmol) and mesylchloride (0.046 g, 0.403 mmol) were added at 0° C. and stirred it atroom temperature for 2h. After completion, reaction was quenched withice-water. Extraction was carried out using DCM; the combined organiclayers were washed with water, brine and dried over anhydrous Na₂SO₄;filtered and concentrated under reduced pressure to afford crudematerial which was purified by prep. HPLC to give the desired compound.LCMS: 582.30 (M+1)⁺; HPLC: 92.74% (@ 254 nm) (R_(t); 5.006); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.55 (s, 1H), 9.86 (bs, 1H), 8.87 (t, 1H, J=4.8Hz), 8.34 (s, 1H), 7.90 (s, 1H), 6.93 (s, 1H), 5.90 (s, 1H), 5.23-5.20(m, 1H), 4.39 (d, 2H, J=4.4 Hz), 4.08 (bs, 2H), 3.80 (bs, 2H), 3.72 (d,2H, J=11.60 Hz), 3.30 (bs, 2H), 3.15 (d, 2H, J=15.60 Hz), 2.93 (s, 3H),2.84-2.76 (m, 2H), 2.27 (m, 1H), 2.21 (s, 3H), 2.12 (s, 3H), 1.81-1.73(m, 2H), 1.51 (d, 6H, J=6.8 Hz).

Step 2: Synthesis ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1′-(methylsulfonyl)-[1,4′-bipiperidin]-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide

To a stirred solution ofN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(1-(1-(methylsulfonyl)piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-pyrazolo[3,4-b]pyridine-4-carboxamide(0.06 g, 0.18 mmol) in EtOH (2 mL), 10% Pd/C in catalytic amount wasadded and stirred it at room temperature under hydrogen pressure(balloon pressure) for 3-4 hr. On completion of reaction, reaction masswas filtered through celite bed, then filtrate was concentrated underreduced pressure, finally the residue was washed with diethyl ether toprovide the desired compound (27% yield). LCMS: 584.35 (M+1)⁺; HPLC:96.59% (@ 254 nm) (R_(t); 4.927); ¹H NMR (CD3OD, 400 MHz) δ 8.28 (s,1H), 7.43 (s, 1H), 6.13 (s, 1H), 5.27 (bs, 1H), 4.55 (s, 2H), 3.94 (d,2H, J=10.4 Hz), 5.74 (d, 2H, J=10.8 Hz), 3.40 (m, 2H), 3.30 (m, 2H),2.89 (s, 3H), 2.85 (m, 2H), 2.41-2.25 (m, 12H), 1.90 (m, 2H), 1.56 (d,6H, J=4.4 Hz).

Syntheses of Compounds B-36 through B-148, B-151 through B-156, andB-164

Compounds B-36 through B-148, B-151 through B-156, and B-164 weresynthesized by methods similar to those described for Compounds B-1through B-35 or by reaction schemes depicted in the general schemes.

Synthesis of Compound C-1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide

Step 1: Synthesis of 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one

3-Cyano-2,4-dimethyl-2-hydroxypyridine (0.3 g, 2.0 mmol) was dissolvedin MeOH (5 mL), to which catalytic amount of Raney Ni and of aqueous NH₃(0.3 mL) were added and the reaction mixture was stirred under hydrogenpressure (bladder pressure) for 3-4 h. After completion of the reaction,catalyst was filtered off and the filtrate was concentrated underreduced pressure. The residue was thoroughly dried to provide thedesired product (0.3 g, quantitative yield).

Step 2: 5-bromo-2-methyl-3-nitrobenzoic acid

1,3-dibromo-5,5-dimethyl-2,4-imidazolidinedione (13.0 g, 45.7 mmol) wasadded to a mixture of 2-methyl-3-nitrobenzoic acid (15 g, 82.8 mmol) inconc. H₂SO₄ (60 mL), and the reaction mixture stirred at roomtemperature for 5 h. After completion of reaction, the mixture wasslowly poured onto ice cold water (400 mL). The precipitated wasfiltered and dried under vacuum to obtain desired5-bromo-2-methyl-3-nitrobenzoic acid (21 g, 98.2%).

Step 3: methyl 5-bromo-2-methyl-3-nitrobenzoate

To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid (16 g,61.5 mmol) in DMF (160 mL), was added iodomethane (35.7 g, 248 mmol) andsodium carbonate (26.3 g, 248 mmol). The resulting reaction mixture wasstirred at 60° C. for 8 h. On completion, the reaction mixture wasfiltered and the inorganic solid residue washed with ethyl acetate. Thecombined filtrates were concentrated under vacuum till dry andre-dissolved in ethyl acetate before washing with 5% sodium bicarbonatesolution (700 mL) followed by 5M HCl solution (300 mL). The organiclayer was finally washed with brine, dried over sodium sulfate andconcentrated to afford pure methyl 5-bromo-2-methyl-3-nitrobenzoate (16g, 94.5%).

Step 4: methyl 3-amino-5-bromo-2-methylbenzoate

A mixture of methyl 5-bromo-2-methyl-3-nitrobenzoate (17 g, 62.0 mmol)in ethanol (85 mL) had NH₄Cl solution (17 g in 85 mL water, 317.8 mmol)followed by Fe powder (27.8 g, 498.1 mmol) added. The resulting reactionmixture was stirred at 90° C. for 1 h. On completion, the reactionmixture was filtered and the filtrate was concentrated till dry. Theresulting solid was dissolved in sat. sodium bicarbonate solution andextracted with ethyl acetate. The combined organic layers were driedover sodium sulfate and concentrated to afford solid methyl3-amino-5-bromo-2-methylbenzoate (15 g, 99.1%).

Step 5: methyl 1-acetyl-6-bromo-1H-indazole-4-carboxylate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (15 g,61.5 mmol) in chloroform (150 mL), was added potassium acetate (6.32 g,64.4 mmol) and acetic anhydride (12.6 g, 122.9 mmol) and reactionmixture was stirred at room temperature for 12 h. After this time,tert-butyl nitrite (25.3 g, 246.1 mmol) and 18-crown-6 (5.7 g, 21.5mmol) were added and reaction stirred again at 65° C. for 3 h. Oncompletion, the reaction mass was cooled to room temperature, dilutedwith chloroform (500 mL) and washed with sat. sodium bicarbonatesolution. The organic layer was dried over sodium sulfate andconcentrated to afford the title compound (18 g, 98.3%).

Step 6: methyl 6-bromo-1H-indazole-4-carboxylate

To a stirred solution of methyl1-acetyl-6-bromo-1H-indazole-4-carboxylate (18 g, 61.0 mmol) in methanol(350 mL), 6N HCl (350 mL) was added and stirred it at 60° C. for 8 h. Oncompletion of reaction, solvent was removed under reduced pressure thenbasified with saturated NaHCO₃ solution till pH 8. The solid precipitatewas filtered and dried under vacuum before being stirred in diethylether for 15 min, filtered and dried to afford methyl6-bromo-1H-indazole-4-carboxylate (11 g, 71.7%).

Step 7: methyl 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate

To a stirred solution of methyl 6-bromo-1H-indazole-4-carboxylate (10 g,39.3 mmol) in acetonitrile (100 mL), was added cesium carbonate (19.2 g,59.05 mmol) followed by bromocyclopentane (11.93 g, 78.3 mmol). Thereaction mass was stirred at 90° C. for 3-4 h. On completion ofreaction, acetonitrile was removed under reduced pressure and wateradded. Extraction was carried out using ethyl acetate and the combinedorganic layers were washed with water, brine and dried over anhydrousNa₂SO₄. The solvent was removed under reduced pressure and the residuepurified by silica gel column chromatography to obtain methyl6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (3.7 g, 29.2%). Theregiochemistry of the cyclopentyl group was confirmed by a NOEexperiment.

Step 8:6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Aqueous NaOH (0.68 g, 17.18 mmol) was added to a solution of methyl6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (3.7 g, 11.45 mmol) inEtOH (40 mL) and stirred at 60° C. for 1 h. After completion of thereaction, ethanol was removed under reduced pressure and the residueacidified using 1N HCl solution. The resulting precipitate was filteredand dried under vacuum. This crude acid (3.2 g, 10.3 mmol) was thendissolved in DMSO (20 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (3.15 g, 20.77 mmol) addedto it. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (8.1 g, 15.5 mmol) was added and left to stir overnight.After completion of the reaction, reaction mass was poured into ice toobtain solid which was filtered and washed with acetonitrile then etherto provide the desired6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(2.6 g, 56.5%). LCMS: 443.05 (M+1)⁺; HPLC: 95.00% (@ 254 nm) (R_(t);7.195); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.61 (t, 1H, J=4.8Hz), 8.36 (s, 1H), 8.19 (s, 1H), 7.70 (s, 1H), 5.88 (s, 1H), 5.23-5.20(m, 1H), 4.33 (d, 2H, J=4.8 Hz), 2.20 (s, 3H), 2.12 (s, 3H), 1.98 (m,2H), 1.97-1.90 (m, 2H), 1.87-1.86 (m, 2H), 1.73-1.67 (m, 2H).

Step 9:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide

To a stirred solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.25 g, 0.56 mmol),4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine(0.20 g, 0.67 mmol) and Pd(PPh₃)₄ (0.032 g, 0.027 mmol) in 1,4-dioxane(4 mL) and purged with argon for 10 min. 2M Na₂CO₃ solution (0.22 g,2.03 mmol) was then added to it before a further argon purge for 10 min.The reaction mixture was stirred at 100° C. for 3 h. After completion ofthe reaction, water was added and extraction carried out using 10%MeOH/DCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide(68.4% yield). LCMS: 540.25 (M+1)⁺; HPLC: 99.15% (@ 254 nm) (R_(t);5.583); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.51 (bs, 1H), 8.62 (t, 1H, J=4.8Hz), 8.36 (s, 1H), 8.10 (s, 1H), 7.85 (s, 1H), 7.80 (d, 2H, J=8 Hz),7.43 (d, 2H, J=8 Hz), 5.88 (s, 1H), 5.34-5.31 (m, 1H), 4.38 (d, 2H,J=4.8 Hz), 3.59-3.58 (m, 4H), 3.52 (s, 2H), 2.38 (m, 4H), 2.21 (s, 3H),2.11 (s, 3H), 2.04-1.99 (m, 2H), 1.88 (m, 4H), 1.72-1.69 (m, 2H).

Synthesis of Compound C-2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(6-formylpyridin-3-yl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(6-formylpyridin-3-yl)-1H-indazole-4-carboxamide

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.1 g, 0.2 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinaldehyde (0.064 g,0.274 mmol) and Pd(PPh₃)₄ (0.013 g, 0.011 mmol) in 1,4-dioxane (2 mL)was purged with argon for 10 min. A solution of 2M Na₂CO₃ (0.087 g,0.820 mmol) was then added to it and argon again purged for 10 min. Thereaction mixture was stirred at 100° C. for 1 h. After completion of thereaction, water was added and extraction was carried out using ethylacetate. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(6-formylpyridin-3-yl)-1H-indazole-4-carboxamide(0.1 g, 94.3%).

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(6-formylpyridin-3-yl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(6-formylpyridin-3-yl)-1H-indazole-4-carboxamide(0.1 g, 0.212 mmol) in methanol (2 mL) was added acetic acid (0.013 g,0.212 mmol) and morpholine (0.056 g, 0.636 mmol) and stirred it at roomtemperature for 3h. Then NaBH₃CN (0.014 g, 0.212 mmol) was added to itand the reaction mixture stirred at room temperature overnight. Aftercompletion of reaction, solvent was removed under reduced pressure andwater was added to it. Extraction was carried out using 10% MeOH/DCM.The combined organic layers were washed with water, dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to givecorresponding1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(6-(morpholinomethyl)pyridin-3-yl)-1H-indazole-4-carboxamide(0.049 g, 43.4%). LCMS: 541.30 (M+1)⁺; HPLC: 94.30% (@ 254 nm) (R_(t);5.379); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.98 (s, 1H), 8.62(t, 1H, J=4.4& 4.8 Hz), 8.39 (s, 1H), 8.21 (s, 2H), 7.90 (s, 1H), 7.56(d, 1H, J=8.4 Hz), 5.88 (s, 1H), 5.35-5.31 (m, 1H), 4.39 (d, 2H, J=4.8Hz), 3.65 (s, 2H), 3.60 (t, 4H, J=4& 4.4 Hz), 2.44 (bs, 4H), 2.21 (s,3H), 2.15 (bs, 2H), 2.11 (s, 3H), 2.05-1.98 (m, 2H), 1.89 (bs, 2H),1.75-1.65 (m, 2H).

Synthesis of Compound C-3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(hydroxymethyl)phenyl)-1H-indazole-4-carboxamide

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1.5 g, 3.5 mmol), (4-(hydroxymethyl)phenyl)boronic acid (0.65 g, 4.30mmol) and Pd(PPh₃)₄ (0.41 g, 0.35 mmol) in 1,4-dioxane (20 mL) waspurged with argon for 10 min. 2 M Na₂CO₃ solution (1.37 g, 12.91 mmol)was then added to it and argon purged again for 10 min. The reactionmixture was stirred at 100° C. for 2 h. After completion of thereaction, water was added to it and extraction was carried out usingDCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(hydroxymethyl)phenyl)-1H-indazole-4-carboxamide(1.5 g, 89.3%).

Step 2:6-(4-(bromomethyl)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(hydroxymethyl)phenyl)-1H-indazole-4-carboxamide(1.5 g, 3.2 mmol) in DCM (20 mL), triphenyl phosphine (1.33 g, 5.10mmol) was added and stirred at room temperature for 10 min. CBr₄ (1.69g, 5.10 mmol) was then added portion-wise to it and the resultingsolution stirred at room temperature overnight. After completion, waterwas added and extraction was carried out using DCM. The combined organiclayers were washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give6-(4-(bromomethyl)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1.0 g, 58.8%).

Step 3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((dimethylamino)methyl)phenyl)-1H-indazole-4-carboxamide

To a stirred solution of6-(4-(bromomethyl)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.15 g, 0.28 mmol) in DMF (1 mL), dimethyl amine (0.163 g, 1.407 mmol)was added and stirred it at room temperature for overnight. Oncompletion, reaction was quenched with ice cold water and extractedusing 10% MeOH/DCM. The combined organic layers were washed with water,dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to afford crude material which was purified by prep. HPLC toprovide the TFA salt of the target compound (0.77 g, 55.7%). LCMS:498.35 (M+1)⁺; HPLC: 99.95% (@ 254 nm) (R_(t); 5.488; ¹H NMR (DMSO-d₆,400 MHz) δ 11.53 (bs, 1H), 9.80 (bs, 1H), 8.64 (t, 1H, J=4.8 Hz), 8.38(s, 1H), 8.19 (s, 1H), 8.00 (d, 2H, J=7.6 Hz), 7.90 (s, 1H), 7.62 (d,2H, J=8.4 Hz), 5.89 (s, 1H), 5.36-5.31 (m, 1H), 4.39 (d, 2H, J=4.8 Hz),4.35 (d, 2H, J=4.4 Hz), 2.77 (s, 6H), 2.22 (s, 3H), 2.16-2.14 (m, 2H),2.12 (s, 3H), 2.04-1.99 (m, 2H), 1.93-1.89 (m, 2H), 1.73-1.70 (m, 2H).

Synthesis of Compound C-4:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((3-(dimethylamino)propoxy)methyl)phenyl)-1H-indazole-4-carboxamide

To a suspension of 60% NaH (0.0112 g, 0.281 mmol) in THF (5 mL),3-(dimethylamino)propan-1-ol (0.039 g, 0.375 mmol) was added at 0° C.followed by the addition of6-(4-(bromomethyl)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.1 g, 0.187 mmol) and stirred at room temperature for 30 min. Oncompletion of reaction, reaction was quenched with ice cold water andextraction carried out using 10% MeOH/DCM. The combined organic layerswere washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by prep. HPLC to provide the TFA salt of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-((3-(dimethylamino)propoxy)methyl)phenyl)-1H-indazole-4-carboxamide.LCMS: 556.35 (M+1)⁺; HPLC: 99.88% (@ 254 nm) (R_(t); 5.470); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 8.65 (t, 1H), 8.39 (s, 1H), 8.22 (s,1H), 8.02 (d, 2H, J=8 Hz), 7.92 (s, 1H), 7.68 (d, 2H, J=8 Hz), 5.89 (s,1H), 5.37-5.33 (m, 1H), 4.59 (s, 2H), 4.39 (d, 2H, J=4.8 Hz), 3.39-3.38(m, 4H), 3.00 (s, 6H), 2.35 (m, 2H), 2.22 (s, 3H), 2.14-2.12 (m, 2H),2.12 (s, 3H), 2.04-2.00 (m, 2H), 1.90 (m, 2H), 1.71-1.70 (m, 2H).

Synthesis of Compound C-5:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide

Step 1: methyl 6-bromo-1-isopropyl-1H-indazole-4-carboxylate

To a stirred solution of methyl 6-bromo-1H-indazole-4-carboxylate (4 g,14.92 mmol) in acetonitrile (30 mL), cesium carbonate (9.7 g, 29.8 mmol)was added followed by 2-iodopropane (3.8 g, 22.3 mmol) and the reactionmixture stirred at 90° C. for 2 h. On completion, the reaction mixturewas concentrated under reduced pressure and the residue diluted withwater and extracted with ethyl acetate. The combined organic layers werewashed with water, brine and dried over sodium sulfate. Solvent wasremoved under reduced pressure to afford crude material which waspurified by column chromatography to afford methyl6-bromo-1-isopropyl-1H-indazole-4-carboxylate (1.5 g, 32.6%).

Step 2:6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-indazole-4-carboxamide

Aqueous NaOH (1.5 equiv in 1 mL water) was added to a solution of methyl6-bromo-1-isopropyl-1H-indazole-4-carboxylate (1.5 g, 4.8 mmol) in EtOH(20 mL) and stirred at 60° C. for 1 h. After completion of the reaction,ethanol was removed under reduced pressure and acidified using 1N HClsolution. Extraction was carried out using ethyl acetate and thecombined organic layers washed with water, brine and dried overanhydrous Na₂SO₄ before filtration and concentrated under reducedpressure. The crude acid (1.26 g, 4.45 mmol) was then dissolved in DMSO(5 mL) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (1.35 g, 8.90mmol) was added. The reaction mixture was stirred at room temperaturefor 15 min before PYBOP (3.47 g, 6.67 mmol) was added to it and stirringcontinued overnight. After completion of the reaction, the mixture waspoured into ice and the resulting precipitate filtered and washed withacetonitrile followed by ether to provide6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-indazole-4-carboxamide(0.8 g, 43.2%).

Step 3:N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide

A solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-1H-indazole-4-carboxamide(0.30 g, 0.72 mmol),4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)morpholine(0.26 g, 0.865 mmol) and Pd(PPh₃)₄ (0.042 g, 0.036 mmol) in 1,4-dioxane(6 mL) was purged with argon for 10 min. 2M Na₂CO₃ solution (0.27 g,2.54 mmol) was then added before an additional argon purged for 10 min.The reaction mixture was stirred at 80° C. for 2 h. After completion ofthe reaction, water was added to it and extraction was carried out usingEtOAc. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive correspondingN-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-isopropyl-6-(4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide(0.20 g, 55.7%). LCMS: 514.25 (M+1)⁺; HPLC: 99.71% (@ 254 nm) (R_(t);5.075); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 8.63 (t, 1H), 8.37(s, 1H), 8.09 (s, 1H), 7.85 (s, 1H), 7.81 (d, 2H, J=6.8 Hz), 7.43 (d,2H, J=7.6 Hz), 5.88 (s, 1H), 5.17-5.14 (m, 1H), 4.38 (d, 2H, J=3.2 Hz),3.59 (m, 4H), 3.52 (s, 2H), 2.38 (m, 4H), 2.21 (s, 3H), 2.12 (s, 3H),1.49 (d, 6H, J=6.8 Hz).

Synthesis of Compound C-6:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2-methyl-4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-formyl-2-methylphenyl)-1H-indazole-4-carboxamide

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.2 g, 0.45 mmol),3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde(0.138 g, 0.56 mmol) and Pd(PPh₃)₄ (0.052 g, 0.045 mmol) in 1,4-dioxane(6 mL) was purged with argon for 10 min. 2 M Na₂CO₃ solution (0.14 g,1.35 mmol) was then added to it and mixture purged with argon again for10 min. The reaction mixture was stirred at 100° C. for 2 h. Aftercompletion of the reaction, water was added to it and the extraction wascarried out using 5% MeOH in DCM. The combined organic layers werewashed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-formyl-2-methylphenyl)-1H-indazole-4-carboxamide(0.15 g, 69%).

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2-methyl-4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-formyl-2-methylphenyl)-1H-indazole-4-carboxamide(0.15 g, 0.31 mmol) in methanol (5 mL), morpholine (0.081 g, 0.93 mmol)and acetic acid (0.01 g, 0.311 mmol) were added at room temperature andreaction stirred at room temperature for 18 h. Then NaBH₃CN (0.023 g,0.37 mmol) was added and reaction stirred again at room temperature for18 h. On completion, methanol was removed under reduced pressure andwater added to the residue. The aqueous phase was extracted with 10%MeOH/DCM and the combined organic layers dried over sodium sulfate andconcentrated to obtain crude material which was purified by columnchromatography to afford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2-methyl-4-(morpholinomethyl)phenyl)-1H-indazole-4-carboxamide(34% yield). LCMS: 554.30 (M+1)⁺; HPLC: 95.54% (@ 254 nm) (R_(t);5.595); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.49 (s, 1H), 8.52 (t, 1H), 8.38(s, 1H), 7.78 (s, 1H), 7.54 (s, 1H), 7.02-7.28 (m, 3H), 5.86 (s, 1H),5.22-5.26 (m, 1H), 4.34 (d, 2H, J=4.4 Hz), 3.58 (t, 4H), 3.47 (s, 2H),2.37 (s, 4H), 2.25 (s, 3H), 2.18 (s, 3H), 2.09-2.06 (m, 2H), 2.09 (s,3H), 1.98-2.03 (m, 2H), 1.86 (m, 2H), 1.66-1.69 (m, 2H).

Synthesis of Compound C-7:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-(((2-(methylsulfonyl)ethyl)amino)methyl)furan-2-yl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-6-(5-formylfuran-2yl)-1H-indazole-4-carboxamide

To a stirred solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.2 g, 0.45 mmol) and (5-formylfuran-2yl)boronic acid (0.075 g, 0.542mmol), in dioxane/water mixture (4 mL+1 mL), Cs₂CO₃ (0.368 g, 1.13 mmol)was added and solution purged with argon for 15 min. Pd(PPh₃)₄ (0.052 g,0.045 mmol) was then added and argon was purged again for 10 min. Thereaction mixture was heated at 100° C. for 2 h. On completion, thereaction mixture was diluted with water and extracted with 10% MeOH/DCM.The combined organic layers were dried over Na₂SO₄ and solvent removedunder reduced pressure to afford crude material which was purified bycolumn chromatography to afford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-6-(5-formylfuran-2yl)-1H-indazole-4-carboxamide(0.09 g, 43.4%).

Step 2: 2-(methylsulfonyl)ethanamine

To a stirred solution of 2-(methylsulfonyl)acetonitrile (1 g, 8.40 mmol)in dry THF (20 mL), borane DMS (1.2 g, 16.8 mmol) was added undercooling condition very slowly. Resulting reaction mixture was heated at40° C. for 2 h and then at room temperature for 12 h. After 12 h MeOH(20 mL) was added at 0° C. very slowly and reaction mixture was refluxedat 80° C. for 1 h. MeOH was removed using reduced pressure and againMeOH (5 mL) was added and heated for 1 h. After 1 h MeOH was removedunder reduced pressure and dioxane in HCl was added and heated for 1 hat 100° C. On completion, the reaction mixture was cooled and DCM addedwith the resulting precipitate filtered and washed with 10% MeOH/DCM,giving a white solid which was dried under vacuum to yield2-(methylsulfonyl)ethanamine (0.95 g, 90%).

Step 3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-(((2-(methylsulfonyl)ethyl)amino)methyl)furan-2-yl)-1H-indazole-4-carboxamide

To a stirred solution1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-methyl)-6-(5-formylfuran-2yl)-1H-indazole-4-carboxamide(0.09 g, 0.19 mmol) in methanol (2 mL), 2-(methylsulfonyl)ethanamine(0.072 g, 0.581 mmol) and DIPEA (0.05 g, 0.39 mmol) was added at roomtemperature and reaction stirred for 3 h. Then NaBH₃CN (0.012 g, 0.196mmol) was added and reaction stirred at room temperature for 18 h. Oncompletion, methanol was removed under reduced pressure and then waterwas added to the residue. The aqueous phase was extracted with 10%MeOH/DCM and the combined organic layers were dried over sodium sulfateand concentrated to obtain crude material which was purified by columnchromatography to afford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-(((2-(methylsulfonyl)ethyl)amino)methyl)furan-2-yl)-1H-indazole-4-carboxamide.LCMS: 566.90 (M+1)⁺; HPLC: 90.05% (@ 254 nm) (R_(t); 5.253); ¹H NMR(MeOD, 400 MHz) δ 8.31 (s, 1H), 8.07 (s, 1H), 7.87 (s, 1H), 6.92 (s,1H), 6.44 (s, 1H), 6.13 (s, 1H), 5.18-5.42 (m, 1H), 4.59 (s, 2H), 4.57(s, 2H), 3.91 (s, 2H), 3.16 (t, 2H, J=6.4 Hz), 3.02 (s, 3H), 2.42 (s,3H), 2.25 (s, 3H), 2.19-2.23 (m, 2H), 2.11-2.14 (m, 2H), 1.98 (m, 2H),1.80 (m, 2H).

Synthesis of Compounds C-8 and C-9:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1H-indazole-4-carboxamideand1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(((2-hydroxyethyl)amino)methyl)phenyl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1H-indazole-4-carboxamide

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1.5 g, 3.4 mmol), 3-hydroxyphenyl boronic acid (0.614 g, 4.07 mmol) andPd(PPh₃)₄ (0.391 g, 0.33 mmol) in 1,4-dioxane (20 mL) was purged withargon for 10 min. Then, 2 M Na₂CO₃ solution (1.29 g, 12.21 mmol) wasadded to it and argon was purged again for 10 min. The reaction mixturewas stirred at 80° C. for 2 h. After completion of the reaction, waterwas added to it and extraction carried out using DCM. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography over silica gel(60-120 mesh size) to afford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1H-indazole-4-carboxamide(1.2 g, 75.4%). LCMS: 471.20 (M+1)⁺; HPLC: 97.18% (@ 254 nm) (R_(t);6.614); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.51 (s, 1H), 8.63 (t, 1H, J=4.8Hz), 8.35 (s, 1H), 8.08 (s, 1H), 7.84 (s, 1H), 7.77 (s, 1H), 7.71 (d,1H, J=7.6 Hz), 7.45 (t, 1H, J=7.6 Hz), 7.36 (d, 1H, J=7.2 Hz), 5.88 (s,1H), 5.35-5.26 (m, 1H), 5.25 (t, 1H, J=4.4 Hz), 4.60 (d, 2H, J=6 Hz),4.39 (d, 2H, J=4.4 Hz), 2.21 (s, 3H), 2.18-2.12 (m, 2H), 2.11 (s, 3H),2.05-1.97 (m, 2H), 1.92-1.86 (m, 2H), 1.74-1.68 (m, 2H).

Step 2:6-(3-(bromomethyl)phenyl)-1-cyclopentyl-N-((dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-yl)methyl)-6-(3-(hydroxymethyl)phenyl)-1H-indazole-4-carboxamide(1.2 g, 2.6 mmol) in DCM (15 mL), triphenyl phosphine (2 g, 7.7 mmol)was added and reaction mixture stirred at room temperature for 10 min.Finally CBr₄ (2.53 g, 7.65 mmol) was added portion-wise and resultingsolution was stirred at room temperature for 18 h. On completion, waterwas added to the reaction mixture and aqueous layer extracted with 10%MeOH/DCM. Combined organic layer was dried over sodium sulfate andconcentrated to give crude material which was purified by columnchromatography over silica gel (60-120 mesh size) to afford6-(3-(bromomethyl)phenyl)-1-cyclopentyl-N-((dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1.0 g, 74.1%).

Step 3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(((2-hydroxyethyl)amino)methyl)phenyl)-1H-indazole-4-carboxamide

6-(3-(bromomethyl)phenyl)-1-cyclopentyl-N-((dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamidewas reacted with 2-aminoethanol using a general procedure for aminationof benzyl bromides to afford the target compound as a TFA salt (19.3%yield). LCMS: 514.20 (M+1)⁺; HPLC: 98.23% (@ 254 nm) (R_(t); 5.465); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 8.91 (bs, 2H), 8.58 (t, 1H,J=4.8 Hz), 8.35 (s, 1H), 8.11 (s, 1H), 7.98 (s, 1H), 7.78 (d, 1H, J=7.2Hz), 7.87 (s, 1H), 7.52-7.58 (m, 2H), 5.89 (s, 1H), 5.26-5.32 (m, 2H),4.38 (d, 2H, J=4.4 Hz), 4.27 (t, 2H), 3.67 (t, 2H, J=4.8 Hz), 3.02 (m,2H), 2.23 (s, 3H), 2.15-2.18 (m, 2H), 2.12 (s, 3H), 1.90-2.08 (m, 4H),1.70-1.73 (m, 2H).

Synthesis of Compound C-10:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-(methylamino)propoxy)phenyl)-1H-indazole-4-carboxamide

Step 1: Synthesis of methyl1-cyclopentyl-6-(3-hydroxyphenyl)-1H-indazole-4-carboxylate

A solution of methyl 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (1g, 3.93 mmol), (3-hydroxyphenyl)-boronic acid (1.43 g, 4.72 mmol) andPd(PPh₃)₄ (0.227 g, 0.196 mmol) in 1,4-dioxane (10.5 mL) was purged withargon for 10 min. Then, 2 M Na₂CO₃ solution (1.5 g, 14.2 mmol) was addedto it and again argon was purged through it for 10 min. The reactionmixture was stirred at 100° C. for 1 h. After completion of thereaction, water was added to it and extraction was carried out using 5%MeOH/DCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography oversilica gel (60-120 mesh size) to afford the title compound (0.8 g, 61%).

Step 2: methyl1-cyclopentyl-6-(3-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxylate

To a stirred solution of methyl1-cyclopentyl-6-(3-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxylate(0.30 g, 0.89 mmol) in DMF (5 mL), 3-bromopropane-1-ol (0.37 g, 2.67mmol) and K₂CO₃ (0.184 g, 1.33 mmol) were added and the resultingreaction mixture heated at 100° C. for 18 h. On completion, water wasadded to reaction mixture and extracted with 10% MeOH/DCM. Combinedorganic layers were dried and concentrated under reduced pressure givingcrude material which was purified by column chromatography over silicagel to afford methyl1-cyclopentyl-6-(3-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxylate(0.2 g, 57%).

Step 3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxamide

Aqueous NaOH (0.06 g, 1.52 mmol) was added to a solution methyl1-cyclopentyl-6-(3-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxylate(0.4 g, 1.0 mmol) in EtOH (4 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand acidified using dilute HCl up to pH 6 and then with citric acid topH 4. The solid precipitate was filtered and dried under vacuum toafford desired acid (0.35 g, 90%). The acid (0.35 g, 0.923 mmol) wasthen dissolved in DMSO (5 mL) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.28 g, 1.84 mmol) was added to it. Thereaction mixture was stirred at room temperature for 15 min before PyBOP(0.72 g, 1.38 mmol) was added to it and stirring was continued forovernight. On completion, reaction mixture was poured into ice andextracted with 10% MeOH/DCM. Combined organic layers were dried andconcentrated under reduced pressure giving crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxamide(0.35 g, 73%).

Step 4: 6-(3-(3-bromopropoxy)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxamide(0.35 g, 0.68 mmol) in DCM (5 mL), triphenyl phosphine (0.285 g, 1.08mmol) was added and the reaction mixture stirred at room temperature for10 min. Finally CBr₄ (0.36 g, 1.08 mmol) was added portion wise andresulting solution was stirred at room temperature for 4 h. Oncompletion, water was added to the reaction mixture and extracted with5% MeOH/DCM. The combined organic layer was dried over sodium sulfateand concentrated to give crude material which was purified by columnchromatography over silica gel to afford 6-(3-(3-bromopropoxy)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.28 g, 71%).

Step 5:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-(methylamino)propoxy)phenyl)-1H-indazole-4-carboxamide

To a stirred solution of 6-(3-(3-bromopropoxy)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1 equiv.) in THF, methylamine (3 equiv. in THF) was added and resultingreaction mass was heated at 60° C. in a sealed flask. On completion,solvent was removed under reduced pressure and crude material obtainedwas purified by preparative HPLC affording1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-(methylamino)propoxy)phenyl)-1H-indazole-4-carboxamideas the TFA salt (22% yield). LCMS: 528.29 (M+1)⁺; HPLC: 97.33% (@ 254nm) (R_(t); 5.863); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.51 (s, 1H), 8.63 (t,1H), 8.35-8.37 (m, 3H), 8.10 (s, 1H), 7.82 (s, 1H), 7.38-7.46 (m, 3H),6.99 (d, 1H, J=7.2 Hz), 5.88 (s, 1H), 5.30-5.37 (m, 1H), 4.37 (d, 2H,J=4.8 Hz), 4.16 (t, 2H, J=6 Hz), 3.06-3.19 (m, 3H), 2.61 (t, 3H, J=5.2Hz), 2.22 (s, 3H), 2.13-2.17 (m, 2H), 2.11 (s, 3H), 1.97-2.09 (m, 3H),1.84-1.89 (m, 2H), 1.69-1.72 (m, 2H).

Synthesis of Compound C-11:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-(dimethylamino)propoxy)phenyl)-1H-indazole-4-carboxamide

To a stirred solution of 6-(3-(3-bromopropoxy)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1 equiv.) in THF, dimethylamine (3 equiv. in THF) was added andresulting reaction mass was heated at 60° C. in a sealed flask. Oncompletion, solvent was removed under reduced pressure and crudematerial obtained was purified by preparative HPLC affording1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(3-(3-(methylamino)propoxy)phenyl)-1H-indazole-4-carboxamideas the TFA salt (23% yield).

LCMS: 542.25 (M+1)⁺; HPLC: 98.61% (@ 254 nm) (R_(t); 5.936); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.51 (s, 1H), 9.38 (bs, 1H), 8.62 (t, 1H, J=4.4Hz), 8.35 (s, 1H), 8.10 (s, 1H), 7.82 (s, 1H), 7.38-7.46 (m, 3H), 6.99(d, 1H, J=7.6 Hz), 5.88 (s, 1H), 5.30-5.37 (m, 1H), 4.37 (d, 2H, J=4.4Hz), 4.15 (t, 2H, J=6 Hz), 3.23-3.28 (m, 2H), 2.83 (d, 6H, J=4.8 Hz),2.22 (s, 3H), 2.13-2.16 (m, 4H), 2.11 (s, 3H), 1.89-2.05 (m, 4H),1.69-1.72 (m, 2H).

Synthesis of Compound C-12:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(3-(methylamino)propoxy)phenyl)-1H-indazole-4-carboxamide

Step 1: methyl1-cyclopentyl-6-(4-hydroxyphenyl)-1H-indazole-4-carboxylate

A solution of methyl 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate(1.5 g, 4.6 mmol), (4-hydroxyphenyl)-boronic acid (0.768 g, 5.56 mmol)and Pd(PPh₃)₄ (0.536 g, 0.464 mmol) in 1,4-dioxane (15 mL) was purgedwith argon for 10 min. Then, 2 M Na₂CO₃ solution (1.77 g, 16.7 mmol) wasadded to it and again argon was purged through it for 10 min. Thereaction mixture was stirred at 100° C. for 2 h. After completion of thereaction, water was added to it and extraction was carried out using 5%MeOH in DCM. The combined organic layers were washed with water, driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto afford crude material which was purified by column chromatographyover silica gel to afford methyl1-cyclopentyl-6-(4-hydroxyphenyl)-1H-indazole-4-carboxylate (1.1 g,71%).

Step 2: methyl1-cyclopentyl-6-(4-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxylate

To a stirred solution of methyl1-cyclopentyl-6-(4-hydroxyphenyl)-1H-indazole-4-carboxylate (0.5 g, 1.48mmol) in DMF (15 mL), 3-bromopropane-1-ol (0.616 g, 4.46 mmol) and K₂CO₃(0.306 g, 2.23 mmol) were added, resulting reaction mass was heated at100° C. for 4 h. On completion, water was added to reaction mass andextracted with ethyl acetate. Combined organic layers were dried andconcentrated under reduced pressure giving crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford methyl1-cyclopentyl-6-(4-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxylate(0.4 g, 68.2%).

Step 3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxamide

Aqueous NaOH (0.06 g, 1.52 mmol) was added to a solution of methyl1-cyclopentyl-6-(4-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxylate(0.4 g, 1.01 mmol) in EtOH (4 mL) and stirred at 60° C. for 1 h. Aftercompletion of the reaction, ethanol was removed under reduced pressureand acidified using dilute HCl up to pH 6 and then with citric acid topH 4. The solid precipitate was filtered and dried under vacuum toafford desired acid (0.23 g, 59.57%). The acid (0.23 g, 0.61 mmol) wasthen dissolved in DMSO (5 mL) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (0.183 g, 1.21 mmol) was added to it. Thereaction mixture was stirred at room temperature for 15 min before PyBOP(0.472 g, 0.907 mmol) was added to it and stirring was continued forovernight. After completion, reaction mass was poured into ice to andextracted with 10% MeOH/DCM. Combined organic layers were dried andconcentrated under reduced pressure giving crude material which waspurified by column chromatography over silica gel (60-120 mesh size) toafford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxamide(0.23 g, 87.8%).

Step 4:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(3-(methylamino)propoxy)phenyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(3-hydroxypropoxy)phenyl)-1H-indazole-4-carboxamide(0.115 g, 0.265 mmol) in DCM (5 mL), triphenyl phosphine (0.137 g, 0.525mmol) was added and reaction stirred at room temperature for 10 min.Finally CBr₄ (0.173 g, 0.525 mmol) was added portion wise to it andresulting solution was stirred at room temperature for 4 h. Oncompletion, methyl amine (2M solution in THF, 1.32 mmol) was added andreaction stirred at room temperature for 18 h and then heated at 60° C.for 12 h. On completion, solvents were removed under reduced pressuregiving crude material which was purified by preparative HPLC to afford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(3-(methylamino)propoxy)phenyl)-1H-indazole-4-carboxamideas the TFA salt (16.9% yield). LCMS: 528.95 (M+1)⁺; HPLC: 95.55% (@ 254nm) (R_(t); 5.670); NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.60 (t, 1H,J=5.2 Hz), 8.32-8.35 (m, 3H), 8.05 (s, 1H), 7.81-7.84 (m, 3H), 7.06 (d,2H, J=8.4 Hz), 5.89 (s, 1H), 5.29-5.33 (m, 1H), 4.37 (d, 2H, J=4.8 Hz),4.12 (t, 2H, J=6 Hz), 3.07-3.12 (m, 2H), 2.60-2.67 m, 3H), 2.21 (s, 3H),2.14-2.20 (m, 2H), 2.12 (s, 3H), 1.97-2.08 (m, 4H), 1.88 (m, 2H),1.69-1.72 (m, 2H).

Synthesis of Compound C-13:(R)-6-(4-((2-carbamoylpyrrolidin-1-yl)methyl)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Step 1: Synthesis of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-formylphenyl)-1H-indazole-4-carboxamide

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.7 g, 1.6 mmol), 4-formyl boronic acid (0.26 g, 1.73 mmol) andPd(PPh₃)₄ (0.182 g, 0.158 mmol) in 1,4-dioxane (10.5 mL) was purged withargon for 10 min. Then, 2 M Na₂CO₃ solution (0.60 g, 5.61 mmol) wasadded to it and again argon was purged through it for 10 min. Thereaction mixture was stirred at 100° C. for 2 h. After completion of thereaction, water was added to it and extraction was carried out using 5%MeOH in DCM. The combined organic layers were washed with water, driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto afford crude material which was purified by column chromatographyover silica gel (60-120 mesh size) to afford Synthesis of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-formylphenyl)-1H-indazole-4-carboxamide(0.68 g, 92.6%).

Step 2:(R)-6-(4-((2-carbamoylpyrrolidin-1-yl)methyl)phenyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-formylphenyl)-1H-indazole-4-carboxamide(0.175 g, 0.373 mmol) in methanol (3 mL) acetic acid (0.022 g, 0.373mmol) and respective amine [[[amine quantity?]]] was added, resultingreaction mass was stirred at room temperature for 4 h. To this reactionmixture sodium cyanoborohydride (0.028 g, 0.448 mmol) was added atcooling condition and reaction mixture was stirred at room temperaturefor 48 h. On completion solvent was removed under reduced pressure andwater was added to it, and then extracted with 10% MeOH/DCM. Combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crude. Thecrude was purified by prep HPLC affording desired compound as TFA salt(12.3% yield). LCMS: 567.30 (M+1)⁺; HPLC: 97.31% (@ 254 nm) (R_(t);5.366); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 9.71 (s, 1H), 8.64(t, 1H, J=4.4 Hz), 8.37 (s, 1H), 8.17 (s, 1H), 7.94-7.97 (m, 3H), 7.87(s, 1H), 7.61-7.67 (m, 3H), 5.89 (s, 1H), 5.31-5.38 (m, 1H), 4.38-4.45(m, 4H), 4.41-4.13 (m, 1H), 3.32 (2H merged in solvent peak), 2.22 (s,3H), 2.14-2.21 (m, 3H), 2.12 (s, 3H), 1.82-2.08 (m, 7H), 1.70-1.72 (m,2H).

Synthesis of Compound C-14:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2-ethoxy-4-((4-ethylpiperazin-1-yl)sulfonyl)phenyl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-4-carboxamide

A solution of methyl 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (1g, 2.2 mmol), bis(pinacolato)diboron (2.78 g, 10.9 mmol) and potassiumacetate (0.64 g, 6.5 mmol) in DMSO (15 mL) was purged with argon for 15min. Then Pd₂Cl₂(dppf). DCM (0.089 g, 0.11 mmol) was added. Resultingreaction mixture was stirred at 80° for 4 h. On completion, reaction wasdiluted with ethyl acetate and washed thoroughly with water, brine anddried over sodium sulfate. Solvent was removed to obtain crude materialwhich was purified by column chromatography over silica gel affording1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-4-carboxamide(0.8 g, 72.7%).

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2-ethoxy-4-((4-ethylpiperazin-1-yl)sulfonyl)phenyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-4-carboxamide(1.2 equiv.) and 1-((4-bromo-3-ethoxyphenyl)sulfonyl)-4-ethylpiperazine(1 equiv.) in dioxane/water mixture (5 mL+1 mL), Na₂CO₃ (3.6 equiv.) wasadded and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.1equiv.) was added and argon was purged again for 10 min. Reactionmixture was heated at 100° C. for 2 h. On completion, reaction mixturewas diluted with water and extracted with 10% MeOH/DCM. Combined organiclayers were dried over Na₂SO₄ and solvent removed under reduced pressureto afford crude material. The crude was purified by prep HPLC affordingdesired compound as TFA salt. LCMS: 661.35 (M+1)⁺; HPLC: 99.73% (@ 254nm) (R_(t); 6.134); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.51 (s, 1H), 9.10 (bs,1H), 8.52 (t, 1H, J=4.8 Hz), 8.35 (s, 1H), 7.99 (s, 1H), 7.80 (d, 1H,J=9.2 Hz), 7.79 (s, 1H), 7.63 (s, 1H), 7.43 (d, 1H, J=8.8 Hz), 5.89 (s,1H), 5.21-5.28 (m, 1H), 4.36 (d, 2H, J=4.4 Hz), 4.20 (q, 2H, J=7.2 Hz),3.81 (d, 2H, J=12.4 Hz), 3.45 (2H merged in solvent peak), 3.12 (d, 6H,J=5.6 Hz), 2.22 (s, 3H), 2.13-2.21 (m, 2H), 2.12 (s, 3H), 2.0-2.07 (m,2H), 1.88-1.95 (m, 2H), 1.69-1.75 (m, 2H), 1.29 (t, 3H, J=6.8 Hz), 1.55(t, 3H, J=7.2 Hz).

Synthesis of Compound C-15:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-(morpholinomethyl)pyridin-2-yl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-formylpyridin-2-yl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole-4-carboxamide(0.15 g, 0.31 mmol) and 6-bromonicotinaldehyde (0.047 g, 0.25 mmol) indioxane/water mixture (2 mL+0.5 mL), Cs₂CO₃ (0.219 g, 0.239 mmol) wasadded and solution purged with argon for 15 min. Then Pd(PPh₃)₄ (0.031g, 0.025 mmol) was added and argon was purged again for 10 min. Reactionmixture was heated at 100° C. for 2 h. On completion, reaction mixturewas diluted with water and extracted with 10% MeOH/DCM. Combined organiclayers were dried over Na₂SO₄ and solvent removed under reduced pressureto afford crude material which was purified by column chromatographyover silica gel to afford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-formylpyridin-2-yl)-1H-indazole-4-carboxamide(0.053 g, 37%).

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-(morpholinomethyl)pyridin-2-yl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-formylpyridin-2-yl)-1H-indazole-4-carboxamide(0.053 g, 0.113 mmol) in methanol (2 mL), morpholine (0.028 g, 0.33mmol) and acetic acid (0.007 g, 0.113 mmol) were added and reactionmixture stirred at room temperature for 2 h. After this time, reactionmixture was cooled to 0° C. and NaBH₃CN (0.007 g, 0.113 mmol) was addedto it. Resulting reaction mixture was stirred again at room temperaturefor 6 h. On completion, methanol was removed under reduced pressure andresidue diluted with water. Aqueous layer was extracted with 20%MeOH/DCM. Combined organic solvent was removed under reduced pressureand residue purified by column chromatography over silica gel affordingtarget1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(5-(morpholinomethyl)pyridin-2-yl)-1H-indazole-4-carboxamideas off white solid (50.8% yield). LCMS: 541.30 (M+1)⁺; HPLC: 99.93% (@254 nm) (R_(t); 5.205); ¹H NMR (MeOD, 400 MHz) δ 7.81 (s, 1H), 7.59 (s,2H), 7.33 (s, 1H), 7.23 (d, 1H, J=8 Hz), 7.12 (d, 1H, J=7.2 Hz), 5.32(s, 1H), 4.44-4.47 (m, 1H), 3.78 (s, 2H), 2.90 (m, 4H), 2.82 (s, 2H),1.71 (m, 4H), 1.61 (s, 3H), 1.44 (s, 3H), 1.35-1.43 (m, 2H), 1.34-1.36(m, 2H), 1.84 (m, 2H), 0.99 (m, 2H).

Synthesis of Compounds C-16 through C-35

Compounds C-16 through C-35 were synthesized by methods similar to thosedescribed for Compounds C-1 through C-15 or by reaction schemes depictedin the general schemes.

Synthesis of Compound D-1:5-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Step 1: Synthesis of 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one

3-Cyano-2,4-dimethyl-2-hydroxypyridine (0.3 g, 2.0 mmol) was dissolvedin MeOH (5 mL), to which catalytic amount of Raney Ni and of aqueous NH₃(0.3 mL) were added and the reaction mixture was stirred under hydrogenpressure (bladder pressure) for 3-4 h. After completion of the reaction,catalyst was filtered off and the filtrate was concentrated underreduced pressure. The residue was thoroughly dried to provide thedesired product (0.3 g, quantitative yield).

Step 2: 5-bromo-2-methyl-3-nitrobenzoic acid

1,3-Dibromo-5,5-dimethyl-2,4-imidazolidinedione (13.0 g, 45.7 mmol) wasadded to a mixture of 2-methyl-3-nitrobenzoic acid (15 g, 82.8 mmol) inconc. H₂SO₄ (60 mL), and the reaction mixture stirred at roomtemperature for 5 h. After completion of reaction, the mixture wasslowly poured onto ice cold water (400 mL). The precipitated wasfiltered and dried under vacuum to obtain desired5-bromo-2-methyl-3-nitrobenzoic acid (21 g, 98.2%).

Step 3: methyl 5-bromo-2-methyl-3-nitrobenzoate

To a stirred solution of 5-bromo-2-methyl-3-nitrobenzoic acid (16 g,61.5 mmol) in DMF (160 mL), was added iodomethane (35.7 g, 248 mmol) andsodium carbonate (26.3 g, 248 mmol). The resulting reaction mixture wasstirred at 60° C. for 8 h. On completion, the reaction mixture wasfiltered and the inorganic solid residue washed with ethyl acetate. Thecombined filtrates were concentrated under vacuum till dry andre-dissolved in ethyl acetate before washing with 5% sodium bicarbonatesolution (700 mL) followed by 5M HCl solution (300 mL). The organiclayer was finally washed with brine, dried over sodium sulfate andconcentrated to afford pure methyl 5-bromo-2-methyl-3-nitrobenzoate (16g, 94.5%).

Step 4: methyl 3-amino-5-bromo-2-methylbenzoate

A mixture of methyl 5-bromo-2-methyl-3-nitrobenzoate (17 g, 62.0 mmol)in ethanol (85 mL) had NH₄Cl solution (17 g in 85 mL water, 317.8 mmol)followed by Fe powder (27.8 g, 498.1 mmol) added. The resulting reactionmixture was stirred at 90° C. for 1 h. On completion, the reactionmixture was filtered and the filtrate was concentrated till dry. Theresulting solid was dissolved in sat. sodium bicarbonate solution andextracted with ethyl acetate. The combined organic layers were driedover sodium sulfate and concentrated to afford solid methyl3-amino-5-bromo-2-methylbenzoate (15 g, 99.1%).

Step 5: methyl 1-acetyl-6-bromo-1H-indazole-4-carboxylate

To a stirred solution of methyl 3-amino-5-bromo-2-methylbenzoate (15 g,61.5 mmol) in chloroform (150 mL), was added potassium acetate (6.32 g,64.4 mmol) and acetic anhydride (12.6 g, 122.9 mmol) and reactionmixture was stirred at room temperature for 12 h. After this time,tert-butyl nitrite (25.3 g, 246.1 mmol) and 18-crown-6 (5.7 g, 21.5mmol) were added and reaction stirred again at 65° C. for 3 h. Oncompletion, the reaction mass was cooled to room temperature, dilutedwith chloroform (500 mL) and washed with sat. sodium bicarbonatesolution. The organic layer was dried over sodium sulfate andconcentrated to afford the title compound (18 g, 98.3%).

Step 6: methyl 6-bromo-1H-indazole-4-carboxylate

To a stirred solution of methyl1-acetyl-6-bromo-1H-indazole-4-carboxylate (18 g, 61.0 mmol) in methanol(350 mL), 6N HCl (350 mL) was added and stirred it at 60° C. for 8 h. Oncompletion of reaction, solvent was removed under reduced pressure thenbasified with saturated NaHCO₃ solution till pH 8. The solid precipitatewas filtered and dried under vacuum before being stirred in diethylether for 15 min, filtered and dried to afford methyl6-bromo-1H-indazole-4-carboxylate (11 g, 71.7%).

Step 7: methyl 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate

To a stirred solution of methyl 6-bromo-1H-indazole-4-carboxylate (10 g,39.3 mmol) in acetonitrile (100 mL), was added cesium carbonate (19.2 g,59.05 mmol) followed by bromocyclopentane (11.93 g, 78.3 mmol). Thereaction mass was stirred at 90° C. for 3-4 h. On completion ofreaction, acetonitrile was removed under reduced pressure and wateradded. Extraction was carried out using ethyl acetate and the combinedorganic layers were washed with water, brine and dried over anhydrousNa₂SO₄. The solvent was removed under reduced pressure and the residuepurified by silica gel column chromatography to obtain methyl6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (3.7 g, 29.2%). Theregiochemistry of the cyclopentyl group was confirmed by a NOEexperiment.

Step 8:6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Aqueous NaOH (0.68 g, 17.18 mmol) was added to a solution of methyl6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (3.7 g, 11.45 mmol) inEtOH (40 mL) and stirred at 60° C. for 1 h. After completion of thereaction, ethanol was removed under reduced pressure and the residueacidified using 1N HCl solution. The resulting precipitate was filteredand dried under vacuum. This crude acid (3.2 g, 10.3 mmol) was thendissolved in DMSO (20 mL) and3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (3.15 g, 20.77 mmol) addedto it. The reaction mixture was stirred at room temperature for 15 minbefore PYBOP (8.1 g, 15.5 mmol) was added and left to stir overnight.After completion of the reaction, reaction mass was poured into ice toobtain solid which was filtered and washed with acetonitrile then etherto provide the desired6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(2.6 g, 56.5%). LCMS: 443.05 (M+1)⁺; HPLC: 95.00% (@ 254 nm) (R_(t);7.195); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.61 (t, 1H, J=4.8Hz), 8.36 (s, 1H), 8.19 (s, 1H), 7.70 (s, 1H), 5.88 (s, 1H), 5.23-5.20(m, 1H), 4.33 (d, 2H, J=4.8 Hz), 2.20 (s, 3H), 2.12 (s, 3H), 1.98 (m,2H), 1.97-1.90 (m, 2H), 1.87-1.86 (m, 2H), 1.73-1.67 (m, 2H).

Synthesis of Compound D-2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide

To a stirred solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.5 g, 1.12 mmol), was added2,2,6,6-tetramethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine(0.36 g, 1.35 mmol) and Pd(PPh₃)₄ (0.066 g, 0.057 mmol) in 1,4-dioxane(10 mL) and the flask purged with argon for 10 min. 2M Na₂CO₃ solution(0.43 g, 4.06 mmol) was then added and the flask again purged with argonfor 10 min. The reaction mixture was stirred at 100° C. for 1 h. Aftercompletion, water was added and extraction carried out using 10%MeOH/DCM. The combined organic layers were washed with water, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure toafford crude material which was purified by column chromatography togive1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide(0.53 g, 77.6%). LCMS: 502.55 (M+1)⁺; HPLC: 99.25% (@ 254 nm) (R_(t);5.528); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (bs, 1H), 8.54 (t, 1H), 8.26(s, 1H), 7.75 (s, 1H), 7.62 (s, 1H), 6.26 (s, 1H), 5.88 (s, 1H),5.29-5.24 (m, 1H), 4.37 (d, 2H, J=5.2 Hz), 2.32 (m, 2H), 2.23 (s, 3H),2.12 (s, 3H), 1.99-1.93 (m, 4H), 1.92-1.88 (m, 2H), 1.72-1.69 (m, 2H),1.23 (s, 6H), 1.17 (s, 6H).

Synthesis of Compound D-3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethylpiperidin-4-yl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethyl-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide(0.15 g, 0.29 mmol) in MeOH (2 mL), 10% Pd/C in catalytic amount wasadded and stirred it at room temperature under a hydrogen balloon for 3hr. On completion of reaction, reaction mixture was filtered through acelite bed and the filtrate concentrated under reduced pressure to give1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(2,2,6,6-tetramethylpiperidin-4-yl)-1H-indazole-4-carboxamide(0.09, 66.4%). LCMS: 504.25 (M+1)⁺; HPLC: 99.77% (@ 254 nm) (R_(t);5.460); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.55 (bs, 1H), 8.43 (t, 1H, J=5.2Hz), 8.26 (s, 1H), 7.70 (s, 1H), 7.50 (s, 1H), 5.89 (s, 1H), 5.22-5.18(m, 1H), 4.36 (d, 2H, J=5.2 Hz), 3.33 (1H merged with DMSO peak), 2.22(s, 3H), 2.12 (s, 3H), 2.09 (m, 2H), 1.99-1.87 (m, 4H), 1.73-1.66 (m,4H), 1.52 (m, 2H), 1.32 (s, 6H), 1.18 (s, 6H).

Synthesis of Compound D-4:6-(1-acetyl-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Step 1: tert-butyl4-(1-cyclopentyl-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate

6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(5 g) was converted to tert-butyl4-(1-cyclopentyl-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate(3.8 g, 61.9%) using the general procedure for Suzuki coupling.

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide

tert-butyl4-(1-cyclopentyl-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate(4 g) was converted to1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide(2.5 g, 76.68%) using general procedure for Boc deprotection. LCMS:446.25 (M+1)⁺; HPLC: 99.35% (@ 254 nm) (R_(t); 5.024); ¹H NMR (DMSO-d₆,400 MHz) δ 11.50 (bs, 1H), 8.54 (t, 1H), 8.28 (s, 1H), 7.75 (s, 1H),7.67 (s, 1H), 6.40 (s, 1H), 5.88 (s, 1H), 5.24 (m, 1H), 4.35 (d, 2H,J=5.2 Hz), 3.25 (m, 2H), 2.93 (t, 2H, J=4.8 & 5.2 Hz), 2.45 (bs, 2H),2.20 (s, 3H), 2.12 (s, 3H), 2.10 (m, 2H), 2.02-1.94 (m, 2H), 1.86 (m,2H), 1.68 (m, 2H).

Step 3:6-(1-acetyl-1,2,3,6-tetrahydropyridin-4-yl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

A stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide(0.1 g, 0.2 mmol) in DCM (3 mL) was cooled to 0° C. Triethyl amine (0.05g, 0.49 mmol) and acetyl chloride (0.02 g, 0.27 mmol) were then added.The reaction mixture was stirred at room temperature for 12 h. Oncompletion, the reaction mixture was concentrated until dryness andwater added to the residue to obtain solid which was filtered washedwith acetonitrile affording a solid which was purified by prep. HPLC toprovide the TFA salt of the target molecule. LCMS: 488.30 (M+1)⁺; HPLC:96.21% (@ 254 nm) (R_(t); 6.266); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.40 (bs,1H), 8.54 (bs, 1H), 8.30 (s, 1H), 7.82 (d, 1H, J=7.2 Hz), 7.69 (s, 1H),6.37 (bs, 1H), 5.88 (s, 1H), 5.24 (m, 1H), 4.36 (d, 2H, J=3.2 Hz), 4.15(d, 2H, J=18.8 Hz), 3.67 (m, 2H), 2.67 (bs, 1H), 2.57 (bs, 1H), 2.20 (s,3H), 2.11 (s, 6H), 2.05 (s, 2H), 1.99 (m, 2H), 1.87 (m, 2H), 1.69 (m,2H).

Synthesis of Compound D-5:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(1-methylpiperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(piperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide

1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamidewas converted to1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(piperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamidethe general PyBOP coupling with1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid and deprotected withgeneral boc deprotection procedure. The compound was purified by prep.To yield HPLC the TFA salt. LCMS: 557.35 (M+1)⁺; HPLC: 98.46% (@ 254 nm)(R_(t); 5.266); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.52-8.50 (m,2H), 8.30 (s, 1H), 8.25 (bs, 1H), 7.82 (d, 1H, J=10 Hz), 7.69 (s, 1H),6.39 (d, 1H, J=12 Hz), 5.88 (s, 1H), 5.25 (m, 1H), 4.36 (d, 2H, J=4.4Hz), 4.28 (bs, 1H), 4.15 (bs, 1H), 3.78-3.70 (m, 2H), 3.31 (d, 2H,J=10.8 Hz), 3.09-2.93 (m, 5H), 2.69 (bs, 1H), 2.59 (bs, 1H), 2.20 (s,3H), 2.12 (s, 3H), 2.09 (m, 2H), 1.99 (m, 2H), 1.90-1.68 (m, 6H).

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(1-methylpiperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide

1-Cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamidewas converted to desired1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(1-methylpiperidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamideusing a general reductive amination methylation procedure. Crudematerial obtained was purified by column chromatography over silica gel(73.5%). LCMS: 571.45 (M+1)⁺; HPLC: 94.84% (@ 254 nm) (R_(t); 5.354); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.53 (t, 1H), 8.30 (s, 1H), 7.82(s, 1H), 7.69 (s, 1H), 6.38 (d, 1H, J=11.6 Hz), 5.88 (s, 1H), 5.24 (m,1H), 4.36 (d, 2H, J=4.4 Hz), 4.24 (bs, 1H), 4.41 (s, 1H), 7.71 (d, 2H,J=4.4 Hz), 2.76 (m, 2H), 2.67 (m, 1H), 2.61 (m, 1H), 2.56 (m, 1H), 2.19(s, 3H), 2.15 (s, 3H), 2.11 (s, 3H), 1.99-1.80 (m, 8H), 1.72-1.59 (m,6H).

Synthesis of Compound D-6:6-([1,4′-bipiperidin]-4-yl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamide

1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamidewas combined with tert-butyl 4-oxopiperidine-1-carboxylate using ageneral reductive amination procedure and product treated to the generalBoc deprotection procedure and purified by prep. HPLC to afford1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamideas the TFA salt. LCMS: 529.40 (M+1)⁺; HPLC: 98.80% (@ 254 nm) (R_(t);4.495); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 10.17 (bs, 1H), 8.78(bs, 1H), 8.54 (m, 2H), 8.32 (s, 1H), 7.92 (s, 1H), 7.73 (s, 1H), 6.46(s, 1H), 5.89 (s, 1H), 5.28 (m, 1H), 4.37 (d, 2H, J=4.8 Hz), 4.00 (m,2H), 3.75 (m, 1H), 2.97 (m, 4H), 2.40-2.25 (m, 2H), 2.21 (s, 3H), 2.12(s, 3H), 2.10 (m, 2H), 2.05-1.65 (m, 12H).

Step 2:6-([1,4′-bipiperidin]-4-yl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

6-([1,4′-bipiperidin]-4-yl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamidewas prepared using a general Pd—C reduction procedure from1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(1-(piperidin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole-4-carboxamideas the TFA salt (50% yield). LCMS: 531.30 (M+1)⁺; HPLC: 79.88% (@ 254nm) (R_(t); 4.560); ¹H NMR (CD3OD, 400 MHz) δ 8.27 (s, 1H), 7.63 (s,1H), 7.45 (s, 1H), 6.13 (s, 1H), 5.15 (m, 1H), 4.55 (s, 2H), 3.80-3.60(m, 5H), 3.25 (m, 2H), 3.20-3.05 (m, 3H), 2.46 (bs, 2H), 2.42 (s, 3H),2.25 (s, 3H), 2.22 (m, 2H), 2.20-2.05 (m, 8H), 1.97 (m, 2H), 1.78 (m,2H).

Synthesis of Compound D-7:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-((4-methylpiperazin-1-yl)methyl)-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-vinyl-1H-indazole-4-carboxamide

A solution of6-bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1.5 g, 3.386 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane(0.625 g, 4.063 mmol) and Pd(PPh₃)₄ (0.392 g, 0.034 mmol) in 1,4-dioxane(15 mL) and purged with argon for 10 min. A solution of 2M Na₂CO₃ (1.29g, 12.18 mmol) was then added to it before a further argon purge for 10min. The reaction mixture was stirred at 100° C. for 1 h. Aftercompletion of the reaction, water was added to it and extraction wascarried out using 10% MeOH/DCM. The combined organic layers were washedwith water, dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to afford crude material which was purified by columnchromatography to give1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-vinyl-1H-indazole-4-carboxamide(0.9 g, 68.2%).

Step 2:1-cyclopentyl-6-(1,2-dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-vinyl-1H-indazole-4-carboxamide(0.9 g, 2.3 mmol) in DCM (10 mL), was added N-methylmorpholine-N-oxide(0.81 g, 6.92 mmol) at 0° C. OsO₄ (0.147 g, 0.576 mmol, 2.5% solution int-BuOH) was then added. The resulting solution was stirred at roomtemperature for 1 h. After completion of reaction, water was added andextraction carried out using 10% MeOH/DCM. The combined organic layerswere washed with water, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to afford crude material which waspurified by column chromatography to give title compound1-cyclopentyl-6-(1,2-dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.6 g, 61.3%).

Step 3:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-6-(1,2-dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.6 g, 1.4 mmol) in 50% THF/Water (16 mL), was added NaIO₄ (0.9 g, 4.2mmol) at 0° C. and stirred at room temperature for 1 h. After completionof reaction, solvent was removed under reduced pressure and water wasadded to it. Extraction was carried out using 10% MeOH/DCM. The combinedorganic layers were washed with water, dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to afford crudematerial which was purified by column chromatography to give1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1H-indazole-4-carboxamide(0.4 g, 72.1%).

Step 4:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-((4-methylpiperazin-1-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-formyl-1H-indazole-4-carboxamide(0.1 g, 0.255 mmol) in methanol (2 mL) was added acetic acid (0.015 g,0.255 mmol) and 1-methylpiperazine (0.077 g, 0.765 mmol) and stirred itat room temperature for 3 h. NaBH₃CN (0.016 g, 0.255 mmol) was thenadded. The resulting reaction mixture was stirred at room temperatureovernight. After completion of reaction, the solvent was removed underreduced pressure and water was added. Extraction was carried out using10% MeOH/DCM. The combined organic layers were washed with water, driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto afford crude material which was purified by column chromatography togive1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-((4-methylpiperazin-1-yl)methyl)-1H-indazole-4-carboxamide(0.027 g, 28.8%). LCMS: 477.25 (M+1)⁺; HPLC: 99.71% (@ 254 nm) (R_(t);4.649); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 8.39 (t, 1H, J=4.8Hz), 8.27 (s, 1H), 7.71 (s, 1H), 7.50 (s, 1H), 5.89 (s, 1H), 5.19-5.14(m, 1H), 4.35 (d, 2H, J=5.2 Hz), 3.58 (s, 2H), 2.39-2.33 (m, 8H), 2.21(s, 3H), 2.12 (s, 3H), 2.07 (s, 3H), 2.00 (m, 2H), 1.99-1.96 (m, 2H),1.91-1.86 (m, 2H), 1.69-1.68 (m, 2H).

Synthesis of Compound D-8:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(hydroxymethyl)-1H-indazole-4-carboxamide

Synthesis of Compound D-8

A stirred solution of1-cyclopentyl-N-4,6-dimethyloxo,2-dihydropyridine-3-yl)methyl)-6-formyl-1H-indazole-4-carboxamide(0.1 g, 0.255 mmol) in methanol (2 mL) was cooled to 0° C. and NaBH₄(0.096 g, 0.257 mmol) was added to it. The resulting reaction mixturewas stirred at room temperature for 3 h. On completion, the reactionmixture was evaporated under reduced pressure and water was added to theresidue. The aqueous phase was extracted with 10% MeOH/DCM and thecombined organic layers dried over Na₂SO₄ and the solvent removed underreduced pressure to obtain crude material which was purified by columnchromatography over silica gel affording desired compound (0.05 g, 50%).LCMS: 395.20 (M+1)⁺; HPLC: 95.70% (@ 254 nm) (R_(t); 5.532); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.36 (t, 1H), 8.27 (s, 1H), 7.74 (s,1H), 7.50 (s, 1H), 5.88 (s, 1H), 5.35 (t, 1H, J=6& 5.2 Hz), 5.16 (m,1H), 4.64 (d, 2H, J=5.6 Hz), 4.35 (d, 2H, J=5.2 Hz), 2.21 (s, 3H), 2.12(s, 3H), 2.10 (m, 2H), 2.01-1.96 (m, 2H), 1.86 (m, 2H), 1.69 (m, 2H).

Synthesis of Compound D-9:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-methyl-1H-indazole-4-carboxamide

Step 1: 6-(bromomethyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(hydroxymethyl)-1H-indazole-4-carboxamide(0.75 g, 1.90 mmol) in DCM (10 mL), triphenyl phosphine (0.998 g, 3.807mmol) was added at room temperature. The mixture was cooled to 0° C.after 5 minutes then carbon tetrabromide (1.26 g, 3.80 mmol) was added.On completion, reaction mixture was evaporated under reduced pressureand the crude residue purified by column chromatography over silica gelto afford desired6-(bromomethyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-yl)methyl)-1H-indazole-4-carboxamide (0.45 g, 51.84%).

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-methyl-1H-indazole-4-carboxamide

6-(bromomethyl)-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine-3-yl)methyl)-1H-indazole-4-carboxamide (0.05 g, 0.13 mmol) wasdissolved in methanol (2 mL) and Pd—C (10%, 0.01 g) was added to it. Theresulting reaction mixture was stirred at room temperature for 1 h. Oncompletion, the reaction mixture was filtered and concentrated to obtainsolid compound which was washed with ether and pentane affording puretarget compound (0.045 g, 91.89%). LCMS: 379.30 (M+1)⁺; HPLC: 98.15% (@254 nm) (R_(t); 6.603); ¹H NMR (CD3OD, 400 MHz) δ 8.28 (s, 1H), 7.63 (s,1H), 7.48 (s, 1H), 6.70 (s, 1H), 5.12 (m, 1H), 4.61 (s, 2H), 2.56 (s,3H), 2.55 (s, 3H), 2.42 (s, 3H), 2.19-2.14 (m, 2H), 2.12-2.04 (m, 2H),2.00-1.92 (m, 2H), 1.82-1.75 (m, 2H).

Synthesis of Compound D-10:6-chloro-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Step 1: 5-chloro-2-methyl-3-nitrobenzoic acid

A stirred solution of 5-chloro-2-methylbenzoic acid (1 mmol) in conc.H₂SO₄ (2 mL) was cooled to −10° C. and nitrating mixture (0.4 mL ofconc. H₂SO_(4+0.1) mL of conc. HNO₃) was added to it in dropwise manner.The reaction mixture was stirred at the same temperature for 20 min thenwarmed to room temperature. The solid precipitate was filtered and driedto obtain 5-chloro-2-methyl-3-nitrobenzoic acid which was used in nextstep without further purification (yield 60-70%).

Step 2: methyl 5-chloro-2-methyl-3-nitrobenzoate

To a stirred solution of 5-chloro-2-methyl-3-nitrobenzoic acid (1equiv.) in DMF (3 ml per 1 mmol), iodomethane (4 equiv.) and sodiumcarbonate (4 equiv.) were added. The resulting reaction mixture wasstirred at 60° C. for 8 h. On completion, the reaction mixture wasfiltered and the inorganic solid residue washed with ethyl acetate. Thecombined filtrate was concentrated under vacuum till dryness. Theresidue was re-dissolved in ethyl acetate and washed with 5% sodiumbicarbonate solution followed by 5M HCl solution. Organic layer wasfinally washed with brine, dried over sodium sulfate and concentrated toafford pure methyl 5-chloro-2-methyl-3-nitrobenzoate (90-95%).

Step 3: methyl 3-amino-5-chloro-2-methylbenzoate

To a stirred solution of methyl 5-chloro-2-methyl-3-nitrobenzoate (1equiv.) in ethanol (2 mL per 1 mmol), was added NH₄Cl solution (5.1equiv., dissolved in water. Volume of water equivalent to volume ofethanol) followed by Fe powder (8 equiv.). The resulting reactionmixture was stirred at 90° C. for 1 h. On completion, reaction mixturewas filtered and the filtrate concentrated until dryness. The resultingsolid which was dissolved in saturated sodium bicarbonate solution andthe aqueous layer extracted with ethyl acetate. Combined organic layerswere dried over sodium sulfate and concentrated to afford solid methyl3-amino-5-chloro-2-methylbenzoate in quantitative yield.

Step 4: methyl 6-chloro-1H-indazole-4-carboxylate

To a stirred solution of methyl 3-amino-5-chloro-2-methylbenzoate (1equiv.) in chloroform (13 mL per 1 mmol), potassium acetate (1.05equiv.) and acetic anhydride (2 equiv.) were added and reaction mixturestirred at room temperature for 12 h. After this time, tert-butylnitrite (4 equiv.) and 18-crown-6 (0.35 equiv.) were added and thereaction stirred again at 65° C. for 3 h. On completion, the reactionmixture was cooled to room temperature, diluted with chloroform (500 mL)and washed with sat. sodium bicarbonate solution. The organic layer wasdried over sodium sulfate and concentrated to afford the title compound(yield 90-95%).

Step 5: methyl 1-acetyl-6-chloro-1H-indazole-4-carboxylate

To a stirred solution of methyl 6-chloro-1H-indazole-4-carboxylate (1equiv.) in methanol (7 mL per 1 mmol), 6N HCl (7 mL per 1 mmol) wasadded and the mixture stirred at 60° C. for 3 h. On completion ofreaction, the solvent was removed under reduced pressure and basifiedwith saturated NaHCO₃ solution till pH 8. The resulting solidprecipitate was filtered and dried under vacuum. Finally, the solid wasstirred in diethyl ether for 15 min, filtered and dried to afford methyl1-acetyl-6-chloro-1H-indazole-4-carboxylate (yield 70-75%).

Step 6: methyl 6-chloro-1-cyclopentyl-1H-indazole-4-carboxylate

To a stirred solution of methyl1-acetyl-6-chloro-1H-indazole-4-carboxylate (1 equiv.) in acetonitrile(3 mL per 1 mmol), cesium carbonate (1.5 equiv.) was added followed bybromocyclopentane (2 equiv.). The reaction mixture was stirred at 90° C.for 3-4 h. On completion of reaction, acetonitrile was removed underreduced pressure and water was added. Extraction was carried out usingethyl acetate and the combined organic layers were washed with water,brine and dried over anhydrous Na₂SO₄. The solvent was then removedunder reduced pressure and residue was purified by silica gel columnchromatography to obtain methyl6-chloro-1-cyclopentyl-1H-indazole-4-carboxylate (yield 30-35%). Thestructure was confirmed as the desired regioisomer by NOE.

Step 7:6-chloro-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of methyl6-chloro-1-cyclopentyl-1H-indazole-4-carboxylate (1 equiv.) in ethanol(3 mL per 1 mmol), aqueous NaOH solution (1.5 equiv in 1 mL water) wasadded and the reaction mixture stirred at 60° C. for 4 h. On completion,ethanol was removed under reduced pressure and the residue acidifiedwith 1N HCl to pH 6. The resulting solid precipitate was filtered,washed with water and dried to obtain pure corresponding acid (yield80-90%). To a solution of this acid (1 equiv.) in DMSO (2 mL for 1mmol), PyBOP (1.5 equiv.) was added and reaction stirred at roomtemperature for 15 min. At this point,3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equiv.) was added andreaction stirred overnight. On completion, water was added and theprecipitate was filtered and washed with water. This solid was thenstirred with acetonitrile for 10 min and filtered again to obtain puretarget molecule6-chloro-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(yield 50-60%). LCMS: 399.10 (M+1)⁺; HPLC: 99.52% (@ 254 nm) (R_(t);7.047); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.59 (s, 1H), 8.59 (t, 1H, J=4.8Hz), 8.35 (s, 1H), 8.04 (s, 1H), 7.59 (d, 1H, J=1.2 Hz), 5.88 (s, 1H),5.17-5.24 (m, 1H), 4.33 (d, 2H, J=4.8 Hz), 2.19 (s, 3H), 2.12 (s, 3H),2.07-2.11 (m, 2H), 1.92-1.98 (m, 2H), 1.84-1.87 (m, 2H), 1.66-1.69 (m,2H).

Synthesis of Compound D-11:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(trifluoromethyl)-1H-indazole-4-carboxamide

Step 1: 2-methyl-3-nitro-5-(trifluoromethyl)benzoic acid

A stirred solution of 2-methyl-5-(trifluoromethyl)benzoic acid (1 mmol)in conc. H₂SO₄ (2 mL) was cooled to −10° C. and a nitrating mixture (0.4mL of conc. H₂SO_(4+0.1) mL of conc. HNO₃) was added to it in dropwisemanner. The reaction mixture was stirred at same temperature for 20 minbefore warming to room temperature. The resulting precipitate wasfiltered off and dried to obtain2-methyl-3-nitro-5-(trifluoromethyl)benzoic acid which was used in nextstep without further purification (yield 60-70%).

Step 2: methyl 2-methyl-3-nitro-5-(trifluoromethyl)benzoate

To a stirred solution of 2-methyl-3-nitro-5-(trifluoromethyl)benzoicacid (1 equiv.) in DMF (3 ml per 1 mmol), iodomethane (4 equiv.) andsodium carbonate (4 equiv.) were added. The resulting reaction mixturewas stirred at 60° C. for 8 h. On completion, reaction mixture wasfiltered and the inorganic solid residue washed with ethyl acetate. Thecombined filtrate was concentrated under vacuum till dryness and theresidue re-dissolved in ethyl acetate before washing with 5% sodiumbicarbonate solution followed by 5M HCl solution. the organic layer wasfinally washed with brine, dried over sodium sulfate and concentrated toafford pure methyl 2-methyl-3-nitro-5-(trifluoromethyl)benzoate (yield90-95%).

Step 3: methyl 3-amino-2-methyl-5-(trifluoromethyl)benzoate

To a stirred solution of methyl2-methyl-3-nitro-5-(trifluoromethyl)benzoate (1 equiv.) in ethanol (2 mLper 1 mmol), was added NH₄Cl solution (5.1 equiv., dissolved in water.Volume of water equivalent to volume of ethanol) followed by Fe powder(8 equiv.). the resulting reaction mixture was stirred at 90° C. for 1h. On completion, the reaction mixture was filtered and the filtrateconcentrated till dryness. The resulting solid was dissolved insaturated sodium bicarbonate solution and the aqueous layer extractedwith ethyl acetate. The combined organic layers were dried over sodiumsulfate and concentrated to afford solid methyl3-amino-2-methyl-5-(trifluoromethyl)benzoate in quantitative yield.

Step 4: methyl 6-(trifluoromethyl)-1H-indazole-4-carboxylate

To a stirred solution of methyl3-amino-2-methyl-5-(trifluoromethyl)benzoate (1 equiv.) in chloroform(13 mL per 1 mmol), potassium acetate (1.05 equiv.) and acetic anhydride(2 equiv.) were added and reaction mixture stirred at room temperaturefor 12 h. After this time, tert-butyl nitrite (4 equiv.) and 18-crown-6(0.35 equiv.) were added and reaction stirred again at 65° C. for 3 h.On completion, the reaction mixture was cooled to room temperature,diluted with chloroform (500 mL) and washed with sat. sodium bicarbonatesolution. the organic layer was dried over sodium sulfate andconcentrated to afford title the compound (yield 90-95%).

Step 5: methyl 1-acetyl-6-(trifluoromethyl)-1H-indazole-4-carboxylate

To a stirred solution of methyl6-(trifluoromethyl)-1H-indazole-4-carboxylate (1 equiv.) in methanol (7mL per 1 mmol), 6N HCl (7 mL per 1 mmol) was added and stirred it at 60°C. for 3 h. On completion of reaction, the solvent was removed underreduced pressure and the residue basified with saturated NaHCO₃ solutiontill pH 8. The resulting precipitate was filtered and dried undervacuum. Finally, the solid was stirred in diethyl ether for 15 min,filtered and dried to afford methyl1-acetyl-6-(trifluoromethyl)-1H-indazole-4-carboxylate (yield 70-75%).

Step 6: methyl1-cyclopentyl-6-(trifluoromethyl)-1H-indazole-4-carboxylate

To a stirred solution of methyl1-acetyl-6-(trifluoromethyl)-1H-indazole-4-carboxylate (1 equiv.) inacetonitrile (3 mL per 1 mmol), cesium carbonate (1.5 equiv.) was addedfollowed by bromocyclopentane (2 equiv.). The reaction mixture wasstirred at 90° C. for 3-4 h. On completion of reaction, acetonitrile wasremoved under reduced pressure and water was added. Extraction wascarried out using ethyl acetate and the combined organic layers washedwith water, brine and dried over anhydrous Na₂SO₄. Solvent was removedunder reduced pressure and residue was purified by silica gel columnchromatography to obtain methyl1-cyclopentyl-6-(trifluoromethyl)-1H-indazole-4-carboxylate (yield30-35%). The structure was confirmed as the desired regioisomer by NOE.

Step 7:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(trifluoromethyl)-1H-indazole-4-carboxamide

To a stirred solution of methyl1-cyclopentyl-6-(trifluoromethyl)-1H-indazole-4-carboxylate (1 equiv.)in ethanol (3 mL for 1 mmol), aqueous NaOH solution (1.5 equiv in 1 mLwater) was added and reaction stirred at 60° C. for 4 h. On completion,ethanol was removed under reduced pressure and residue acidified with 1NHCl to pH 6. Solid that precipitates out was filtered, washed with waterand dried to obtain pure corresponding acid (yield 80-90%). To asolution of this acid (1 equiv.) in DMSO (2 mL for 1 mmol), PyBOP (1.5equiv.) was added and reaction stirred at rt for 15 min. Then3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2 equiv.) was added andreaction stirred overnight. On completion, water was added and solidthat precipitates out was filtered and washed with water. Then thissolid was stirred with acetonitrile for 10 min and filtered again toobtain1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(trifluoromethyl)-1H-indazole-4-carboxamide(yield 50-60%). LCMS: 433.15 (M+1)⁺; HPLC: 99.51% (@ 254 nm) (R_(t);7.326); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.53 (s, 1H), 8.76 (t, 1H), 8.50(s, 1H), 8.38 (s, 1H), 7.85 (s, 1H), 5.88 (s, 1H), 5.38-5.41 (m, 1H),4.35 (d, 2H, J=4.4 Hz), 2.21 (s, 3H), 2.12-2.20 (m, 2H), 2.12 (s, 3H),1.88-2.00 (m, 4H), 1.69-1.71 (m, 2H).

Synthesis of Compound D-12: tert-butyl4-(6-bromo-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-1-yl)piperidine-1-carboxylate

Step 1: tert-butyl 4-hydroxypiperidine-1-carboxylate

To a solution of piperidin-4-ol (25 g, 247 mmol) in DCM (250 mL),triethyl amine (52 mL, 371 mmol) was added and the resulting solutionwas cooled to 0° C. Boc anhydride (64.75 g, 297 mmol) was added to itslowly and the reaction mixture was stirred at room temp overnight.Water was added to reaction and the organic layer separated. The organiclayer was further washed with water, brine and dried over sodiumsulfate. Solvent was removed under reduced pressure to afford desiredtert-butyl 4-hydroxypiperidine-1-carboxylate (50 g, crude).

Step 2: tert-butyl 4-(tosyloxy)piperidine-1-carboxylate

To a solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (50 g,248.7 mmol) in DCM (500 mL), triethyl amine (104 mL, 746 mmol) was addedand resulting solution cooled to 0° C. p-Toluenesolfonyl chloride wasthen added to it portion-wise and the reaction continued at roomtemperature overnight. On completion, water was added to reaction andorganic layer separated. The organic layer was further washed withwater, brine and dried over sodium sulfate. Solvent was removed underreduced pressure to afford crude material which was purified by silicagel chromatography affording desired tert-butyl4-(tosyloxy)piperidine-1-carboxylate (40 g).

Step 3: methyl 6bromo-1-(tert-butoxycarbonyl)piperidine-4-yl)-1H-indazole-4-carboxylate

To a solution of methyl 6-bromo-1H-indazole-4-carboxylate (5 g, 19.6mmol) in DMF (22 mL), cesium carbonate (9.5 gm, 29.4 mmol) andtert-butyl 4-(tosyloxy)piperidine-1-carboxylate (6.95 g, 19.6 mmol) wereadded and resulting solution was heated at 80° C. for 18 h. Oncompletion, water was added to reaction mixture and the aqueous phaseextracted with ethyl acetate. The combined organic layers were furtherwashed with water, brine and dried over sodium sulfate. The solvent wasremoved under reduced pressure to afford crude material which waspurified by silica gel chromatography affording desired methyl 6bromo-1-(tert-butoxycarbonyl)piperidine-4-yl)-1H-indazole-4-carboxylate(2.4 g, 28%).

Step 4: tert-butyl4-(6-bromo-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-1-yl)piperidine-1-carboxylate

To a stirred solution of methyl 6bromo-1-(tert-butoxycarbonyl)piperidine-4-yl)-1H-indazole-4-carboxylate(1 equiv.) in ethanol (3 mL per 1 mmol), aqueous NaOH solution (1.5equiv.) was added and reaction stirred at 60° C. for 4 h. On completion,the ethanol was removed under reduced pressure and residue acidifiedwith 1N HCl to pH 4-5. The resulting precipitate was filtered, washedwith water and dried to obtain pure acid (yield 75-80%). To a stirredsolution of this acid (1 equiv.) in DMSO (1.5 ml for 1 mmol) was addedPyBop (1.5 equiv) and 3-(aminomethyl)-4,6-dimethylpyridin-2(1H)-one (2equiv). The resulting reaction mixture was stirred at room temperaturefor 12 h. Upon completion, water was added to reaction mixture and theresulting precipitate filtered and washed with acetonitrile to obtaintert-butyl4-(6-bromo-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-1-yl)piperidine-1-carboxylate(yield 55-60%). LCMS: 558.15 (M+1)⁺; HPLC: 98.66% (@ 254 nm) (R_(t);7.314); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.61 (t, 1H), 8.37(s, 1H), 8.26 (s, 1H), 7.71 (s, 1H), 5.88 (s, 1H), 4.92 (m, 1H), 4.33(d, 2H, J=4.4 Hz), 4.10-4.07 (m, 2H), 2.96 (bs, 2H), 2.19 (s, 3H), 2.12(s, 3H), 1.90 (bs, 4H), 1.42 (s, 9H).

Synthesis of Compound D-13:6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide

Step 1:6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide

A stirred solution of tert-butyl4-(6-bromo-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-1-yl)piperidine-1-carboxylate(1 mmol) in DCM (5 mL) was cooled to 0° C. and TFA (2 mL) was added toit. The reaction mixture was stirred at room temperature for 1 h. Oncompletion, the reaction was concentrated to dryness and the residuebasified with aqueous sodium bicarbonate till pH 8 and aqueous layerextracted with 20% MeOH/DCM. The combined organic layers were dried oversodium sulfate and concentrated to afford6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide.

Step 2:6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide

To a stirred solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide(1 equiv.) in DMSO (1.5 ml per 1 mmol), triethylamine (3 equiv.), PyBOP(1.5 equiv.) and pivalic acid (2 equiv.) were added and resultingreaction mixture stirred at room temperature for 12 h. On completion,water was added to reaction mixture, the resulting precipitate wasfiltered and washed with acetonitrile to obtain6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide(75% yield). LCMS: 542.15 (M+1)⁺; HPLC: 99.26% (@ 254 nm) (R_(t);6.583); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.50 (s, 1H), 8.60 (t, 1H), 8.37(s, 1H), 8.27 (s, 1H), 7.71 (s, 1H), 5.87 (s, 1H), 5.01 (m, 1H), 4.43(d, 2H, J=12.4 Hz), 4.33 (d, 2H, J=4.4 Hz), 3.01 (t, 2H, J=11.6& 11.2Hz), 2.19 (s, 3H), 2.12 (s, 3H), 2.00-1.85 (m, 4H), 1.23 (s, 9H).

Synthesis of Compound D-14:1-(1-(azetidine-3-carbonyl)piperidin-4-yl)-6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamidewas coupled with 1-(tert-butoxycarbonyl)azetidine-3-carboxylic acidusing the general PyBOP coupling procedure followed by the general Bocdeprotection method. The crude material was purified by prep HPLC toyield1-(1-(azetidine-3-carbonyl)piperidin-4-yl)-6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamideas the TFA salt (13% yield).

LCMS: 541.15 (M+1)⁺; HPLC: 99.01% (@ 254 nm) (R_(t); 4.738); ¹H NMR(DMSO-d₆, 400 MHz) δ 11.50 (s, 1H), 8.61 (t, 1H, J=4.4 Hz), 8.38 (s,1H), 8.27 (s, 1H), 7.73 (s, 1H), 5.88 (s, 1H), 5.00 (m, 1H), 4.55 (d,1H), 4.33 (d, 2H, J=4.4 Hz), 4.15-4.05 (m, 4H), 4.00-3.95 (m, 1H), 3.65(d, 1H) 3.22 (m, 1H), 2.87 (t, 1H), 2.19 (s, 3H), 2.12 (s, 3H),2.00-1.85 (m, 4H).

Synthesis of Compound D-15:1-(1-benzylpiperidin-4-yl)-6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide(1 equiv.) and benzaldehyde (1.5 equiv.) in methanol (5 mL per 0.3mmol), acetic acid (1 equiv.) was added and reaction stirred at roomtemperature for 5 h. Then NaBH₃CN (1 equiv.) was added and the reactionstirred overnight. On completion, the solvent was removed under reducedpressure and the residue purified by column chromatography (27%). LCMS:548.10 (M+1)⁺; HPLC: 99.16% (@ 254 nm) (R_(t); 5.349); ¹H NMR (DMSO-d₆,400 MHz) δ 11.51 (s, 1H), 8.59 (t, 1H, J=4.8 Hz), 8.36 (s, 1H), 8.25 (s,1H), 7.69 (d, 1H, J=1.2 Hz), 7.34 (d, 4H, J=4.4 Hz), 7.27-7.24 (m, 1H),5.87 (s, 1H), 4.70 (m, 1H), 4.33 (d, 2H, J=4.8 Hz), 3.55 (s, 2H), 2.93(d, 2H, J=10 Hz), 2.30-2.20 (m, 2H), 2.19 (s, 3H), 2.12 (s, 3H), 2.09(m, 2H), 1.87 (m, 2H).

Synthesis of Compound D-16:6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(1′-methyl-[1,4′-bipiperidin]-4-yl)-1H-indazole-4-carboxamide

To a stirred solution of6-bromo-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-(piperidin-4-yl)-1H-indazole-4-carboxamide(1 equiv.) and 1-methylpiperidin-4-one (1.5 equiv.) in methanol (5 mLper 0.3 mmol), acetic acid (1 equiv.) was added and reaction stirred atroom temperature for 5 h. Then NaBH₃CN (1 equiv.) was added and thereaction stirred overnight. On completion, the solvent was removed underreduced pressure and the residue purified by column chromatography (24%yield). LCMS: 555.15 (M+1)⁺; HPLC: 92.59% (@ 254 nm) (R_(t); 4.143); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.52 (s, 1H), 8.60 (t, 1H, J=5.2 Hz), 8.35 (s,1H), 8.24 (s, 1H), 7.68 (s, 1H), 5.88 (s, 1H), 4.63-4.68 (m, 1H), 4.32(d, 2H, J=4.8 Hz), 2.97 (d, 2H, J=11.6 Hz), 2.79 (d, 2H, J=10.8 Hz),2.22-2.40 (m, 3H), 2.19 (s, 3H), 2.13 (s, 3H), 2.11 (s, 3H), 1.99-2.08(m, 2H), 1.81-1.88 (m, 4H), 1.69 (d, 2H, J=11.2 Hz), 1.43-1.48 (m, 2H).

Synthesis of Compound D-17:6-bromo-1-cyclopentyl-N-((4-((dimethylamino)methyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

Step 1: tert-butyldimethyl(prop-2-yn-1-yloxy)silane

To a solution of propargyl alcohol (15 g, 267 mmol) in DMF (60 mL),imidazole (21.85 g, 321 mmol) was added at 0° C. followed by the slowaddition of a solution of TBDMS-Cl (47.2 mL, 272 mmol) in DMF (70 mL).Resulting reaction mixture was stirred at 0° C. for 30 min and then atroom temperature for overnight. On completion, water was added to thereaction mixture and the aqueous layer extracted with 20% ethylacetate/hexane. The combined organic layers were dried over sodiumsulfate and concentrated to obtain crude material which was purified bycolumn chromatography over silica gel affordingtert-butyldimethyl(prop-2-yn-1-yloxy)silane (45.5 g, 32.9%).

Step 2: 5-((tert-butyldimethylsilyl)oxy)pent-3-yn-2-one

To a solution of tert-butyldimethyl(prop-2-yn-1-yloxy)silane (15.1 g,88.3%) in THF (300 mL), n-BuLi (67 mL, 105.9 mmol, 1.6 N in hexane) wasadded slowly at −78° C. After the addition was complete, the temperaturewas increased slowly to 10° C. over a period of 2 h. The reactionmixture was cooled again to −78° C. and BF₃. OEt₂ (14 mL, 105.9 mmol)added slowly and reaction mixture stirred at this temperature for 5 min.Acetic anhydride (11 mL, 115.6 mmol) was slowly added and the reactionwas continued while temperature was allowed to rise to room temperatureover a period of 2 h. After this time, reaction mass was quenched with1N NaOH till pH 7-8 and biphasic solution was extracted with ethylacetate. The organic layer was concentrated to obtain crude materialwhich was further purified by column chromatography over silica gel toyield 5-((tert-butyldimethylsilyl)oxy)pent-3-yn-2-one (9.2 g, 52.6%).

Step 3: 4-(((tert-butyldimethylsilyl)oxy)methyl)-2-hydroxy-6-methylnicotinonitrile

To a solution of 5-((tert-butyldimethylsilyl)oxy)pent-3-yn-2-one (9.2 g,43 mmol) in 90% ethanol (185 mL), cyanoacetamide (4.35 g, 51.8 mmol) andpiperidine acetate. were added and reaction heated at 85° C. for 4 h.The piperidine acetate was prepared by the addition of piperidine to asolution of acetic acid (2.2 mL) in water (4.4 mL) till pH 8. Oncompletion, ethanol was evaporated and water was added to residue. Theresulting solid was filtered and washed with 200 mL water before beingwas stirred again with acetonitrile for 10 min and filtered again toobtain4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile(3.82 g, 29.6%).

Step 4:3-(aminomethyl)-4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methylpyridin-2(1H)-one

To a solution of4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methyl-2-oxo-1,2-dihydropyridine-3-carbonitrile(3.8 g, 13.7 mmol) in methanol (200 mL), Raney Ni and ammonia (20 mL)were added and reaction stirred under hydrogen balloon pressure for 15h. On completion, reaction mixture was filtered through a celite bed andthe filtrate concentrated to obtain3-(aminomethyl)-4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methylpyridin-2(1H)-oneas an off white solid (3.2 g, 83.1%).

Step 5: 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylic acid

To a stirred solution of methyl6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (3 g, 9.28 mmol) inethanol (30 mL), aqueous NaOH solution (0.56 g in 3 mL water, 13.93mmol) was added and reaction stirred at 60° C. for 4 h. On completion,ethanol was removed under reduced pressure and residue acidified with 1NHCl to pH 6. The resulting precipitate was filtered, washed with waterand dried to 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylic acid (2.3 g,80.4%).

Step 6:6-bromo-1-cyclopentyl-N-((4-(hydroxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a solution of 6-bromo-1-cyclopentyl-1H-indazole-4-carboxylic acid(2.3 g, 7.5 mmol) in DMSO (30 mL), PyBOP (5.82 g, 11.20 mmol) was addedand reaction stirred at room temperature for 15 min.3-(Aminomethyl)-4-(((tert-butyldimethylsilyl)oxy)methyl)-6-methylpyridin-2(1H)-one(2.52 g, 8.96 mmol) was then added and reaction mixture stirredovernight. On completion, water was added and the resulting precipitatefiltered and washed with water. This solid was then stirred withacetonitrile for 10 min and filtered again to obtain6-bromo-1-cyclopentyl-N-((4-(hydroxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(1.4 g, 40.9%).

Step 7:6-bromo-1-cyclopentyl-N-((4-((dimethylamino)methyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide

To a stirred solution of6-bromo-1-cyclopentyl-N-((4-(hydroxymethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(0.6 g, 1.3 mmol) in DCM (5 mL), triphenylphosphine (0.54 g, 2.06 mmol)was added at 0° C. followed by carbon tetrabromide (0.684 g, 2.262 mmol)at the same temperature. Reaction mixture was stirred at roomtemperature for 2 h. On completion, the reaction mixture was purified bycolumn over silica gel to afford intermediate6-bromo-N-((4-(bromomethyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1-cyclopentyl-1H-indazole-4-carboxamide(0.4 g, 58.8%). To a solution of this intermediate in DMF, respectiveamine (5 equiv.) was added and reaction stirred for overnight at roomtemperature. On completion, reaction mixture was poured into water andextracted with 15% MeOH/DCM. The combined organic layers were dried oversodium sulfate and concentrated to obtain crude material which waspurified by prep HPLC affording6-bromo-1-cyclopentyl-N-((4-((dimethylamino)methyl)-6-methyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamide(19.2% yield). LCMS: 486.05 (M+1)⁺; HPLC: 99.59% (@ 254 nm) (R_(t);5.723); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.65 (s, 1H), 8.83 (s, 1H), 8.36(s, 1H), 8.20 (s, 1H), 7.62 (s, 1H), 6.07 (s, 1H), 5.23-5.18 (m, 1H),4.41 (d, 2H, J=4 Hz), 3.37 (m, 2H), 2.16 (s, 6H), 2.15-2.05 (m, 5H),1.99-1.91 (m, 2H), 1.86 (m, 2H), 1.69 (m, 2H).

Synthesis of Compounds D-18 and D-19:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(piperidine-4-carbonyl)piperazin-1-yl)-1H-indazole-4-carboxamideand1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(piperazin-1-yl)-1H-indazole-4-carboxamide

Step 1: methyl6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-1-cyclopentyl-1H-indazole-4-carboxylate

To a stirred solution of methyl6-bromo-1-cyclopentyl-1H-indazole-4-carboxylate (5 g, 15.527 mmol) inDMSO (25 mL), 1-Boc piperazine (4.3 g, 23.3 mmol), K₂CO₃ (4.28 g, 31.05mmol), CuI (0.295 g, 1.55 mmol) and L-proline (0.357 g, 3.1 mmol) wereadded in given order and resulting reaction mixture heated to 100° C.for 48 h. On completion, the reaction mixture was poured into water andextracted with DCM. The combined organic layers were washed with waterand dried over sodium sulfate. The solvent was removed under reducedpressure to obtain crude residue which was purified by columnchromatography over silica gel affording methyl6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-1-cyclopentyl-1H-indazole-4-carboxylate(2 g, 30%).

Step 2:6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-1-cyclopentyl-1H-indazole-4-carboxylicacid

To a stirred solution of methyl6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-1-cyclopentyl-1H-indazole-4-carboxylate(2 g, 4.67 mmol) in methanol (20 mL), was added aqueous NaOH solution(0.373 g, dissolved in 10 mL water, 9.34 mmol) and the reaction mixturestirred at 60° C. for 4 h. On completion, ethanol was removed underreduced pressure and the residue acidified with 1N HCl to pH 6. Theprecipitate was filtered, washed with water and dried to obtain pure6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-1-cyclopentyl-1H-indazole-4-carboxylicacid (1.7 g, 88%).

Step 3: tert-butyl4-(1-cyclopentyl-4-(((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-6-yl)piperazine-1-carboxylate

To a solution of6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-1-cyclopentyl-1H-indazole-4-carboxylicacid (1.7 g, 4.1 mmol) in DMSO (10 mL), amine (1.248 g, 8.212 mmol) wasadded and the reaction mixture stirred at room temperature for 10 min.PYBOP (3.202 g, 6.159 mmol) was then added and the reaction mixturestirred at room temperature overnight. On completion, the reactionmixture was poured into water and extracted with DCM. The combinedorganic layers were washed with water and dried over sodium sulfate.Solvent was removed under reduced pressure to obtain crude residue whichwas purified by column chromatography over silica gel affording desiredcompound (2 g, 88%).

Step 4: Synthesis of1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(piperazin-1-yl)-1H-indazole-4-carboxamide

tert-Butyl4-(1-cyclopentyl-4-4(4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)carbamoyl)-1H-indazol-6-yl)piperazine-1-carboxylate(0.1 g) was converted to the target molecule using the general Bocdeprotection procedure. The crude reaction mixture was purified by prepHPLC to afford the TFA salt. LCMS: 449.30 (M+1)⁺; HPLC: 99.11% (@ 254nm) (R_(t); 4.824); ¹H NMR (DMSO-d₆, 400 MHz) δ 11.52 (bs, 1H), 8.80(bs, 2H), 8.41 (t, 1H), 8.17 (s, 1H), 7.34 (s, 1H), 7.21 (s, 1H), 5.88(s, 1H), 5.18-5.11 (m, 1H), 4.35 (d, 2H, J=4 Hz), 3.46 (bs, 4H), 3.28(bs, 4H), 2.20 (s, 3H), 2.11 (s, 3H), 2.08-2.05 (m, 2H), 1.98-1.90 (m,2H), 1.86 (m, 2H), 1.67 (m, 2H).

Step 5:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(piperidine-4-carbonyl)piperazin-1-yl)-1H-indazole-4-carboxamide

1-Cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(piperazin-1-yl)-1H-indazole-4-carboxamidewas converted to the target compound in a 58% yield using the by generalPyBOP coupling procedure. Finally, Boc deprotection with the generalprocedure produced a crude residue which was triturated with diethylether to afford pure1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(4-(piperidine-4-carbonyl)piperazin-1-yl)-1H-indazole-4-carboxamideas the TFA salt (65.3% yield).

Synthesis of Compound D-20:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-isopropyl-1H-indazole-4-carboxamide

Step 1:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(prop-1-en-2-yl)-1H-indazole-4-carboxamide

6-Bromo-1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-1H-indazole-4-carboxamidewas converted to desired compound in 70-80% yield using the generalSuzuki coupling procedure.

Step 2:1-cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-isopropyl-1H-indazole-4-carboxamide

1-Cyclopentyl-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-6-(prop-1-en-2-yl)-1H-indazole-4-carboxamidewas reduced using the standard Pd—C hydrogenation conditions andpurified by prep HPLC to afford the target compound as a TFA salt (35%yield). LCMS: 407.25 (M+1)⁺; HPLC: 97.79% (@ 254 nm) (R_(t); 7.285); ¹HNMR (DMSO-d₆, 400 MHz) δ 11.54 (s, 1H), 8.43 (t, 1H), 8.25 (s, 1H), 7.64(s, 1H), 7.48 (s, 1H), 5.89 (s, 1H), 5.18 (m, 1H), 4.36 (d, 2H, J=4 Hz),3.04 (m, 1H), 2.21 (s, 3H), 2.12 (s, 3H), 2.08 (m, 2H), 1.98 (m, 2H),1.86 (m, 2H), 1.69 (m, 2H), 1.28 (d, 6H, J=6.8 Hz).

Syntheses of Compounds D-21 though D-91 and D-93 through D-98

Compounds D-21 though D-91 and D-93 through D-98 were synthesized bymethods similar to those described for Compounds D-1 through D-20 or byreaction schemes depicted in the general schemes.

Syntheses of Compounds E-1, E-2, F-1, and F-2

Compounds E-1, E-2, F-1, and F-2 were synthesized by methods similar tothose described herein.

Example 2 Bioassay Protocol and General Methods

Protocol for Wild-Type and Mutant PRC2 Enzyme Assays

General Materials. S-adenosylmethionine (SAM), S-adenosylhomocyteine(SAH), bicine, KCl, Tween20, dimethylsulfoxide (DMSO) and bovine skingelatin (BSG) were purchased from Sigma-Aldrich at the highest level ofpurity possible. Dithiothreitol (DTT) was purchased from EMD. ³H-SAM waspurchased from American Radiolabeled Chemicals with a specific activityof 80 Ci/mmol. 384-well streptavidin Flashplates were purchased fromPerkinElmer.

Substrates. Peptides representative of human histone H3 residues 21-44containing either an unmodified lysine 27 (H3K27me0) or dimethylatedlysine 27 (H3K27me2) were synthesized with a C-terminal G(K-biotin)linker-affinity tag motif and a C-terminal amide cap by 21^(st) CenturyBiochemicals. The peptides were high-performance liquid chromatography(HPLC) purified to greater than 95% purity and confirmed by liquidchromatography mass spectrometry (LC-MS). The sequences are listedbelow.

H3K27me0: (SEQ ID NO: 1) ATKAARKSAPATGGVKKPHRYRPGGK(biotin)-amideH3K27me2: (SEQ ID NO: 2) ATKAARK(me2)SAPATGGVKKPHRYRPGGK(biotin)-amide

Chicken erythrocyte oligonucleosomes were purified from chicken bloodaccording to established procedures.

Recombinant PRC2 Enzymes. Human PRC2 enzymes were purified as4-component enzyme complexes co-expressed in Spodoptera frugiperda (sf9)cells using a baculovirus expression system. The subunits expressed werewild-type EZH2 (NM_004456) or EZH2 Y641F, N, H, S or C mutants generatedfrom the wild-type EZH2 construct, EED (NM_003797), Suz12 (NM_015355)and RbAp48 (NM_005610). The EED subunit contained an N-terminal FLAG tagthat was used to purify the entire 4-component complex from sf9 celllysates. The purity of the complexes met or exceeded 95% as determinedby SDS-PAGE and Agilent Bioanalyzer analysis. Concentrations of enzymestock concentrations (generally 0.3-1.0 mg/mL) was determined using aBradford assay against a bovine serum albumin (BSA) standard.

General Procedure for PRC2 Enzyme Assays on Peptide Substrates. Theassays were all performed in a buffer consisting of 20 mM bicine(pH=7.6), 0.5 mM DTT, 0.005% BSG and 0.002% Tween20, prepared on the dayof use. Compounds in 100% DMSO (1 μL) were spotted into polypropylene384-well V-bottom plates (Greiner) using a Platemate 2×3 outfitted witha 384-channel pipet head (Thermo). DMSO (1 μL) was added to columns 11,12, 23, 24, rows A-H for the maximum signal control, and SAH, a knownproduct and inhibitor of PRC2 (1 μL) was added to columns 11,12, 23, 24,rows I-P for the minimum signal control. A cocktail (40 μL) containingthe wild-type PRC2 enzyme and H3K27me0 peptide or any of the Y641 mutantenzymes and H3K27me2 peptide was added by Multidrop Combi (Thermo). Thecompounds were allowed to incubate with PRC2 for 30 min at 25° C., thena cocktail (10 μL) containing a mixture of non-radioactive and ³H-SAMwas added to initiate the reaction (final volume=51 μL). In all cases,the final concentrations were as follows: wild-type or mutant PRC2enzyme was 4 nM, SAH in the minimum signal control wells was 1 mM andthe DMSO concentration was 1%. The final concentrations of the rest ofthe components are indicated in Table 7, below. The assays were stoppedby the addition of non-radioactive SAM (10 μL) to a final concentrationof 600 μM, which dilutes the ³H-SAM to a level where its incorporationinto the peptide substrate is no longer detectable. 50 μL of thereaction in the 384-well polypropylene plate was then transferred to a384-well Flashplate and the biotinylated peptides were allowed to bindto the streptavidin surface for at least 1 h before being washed threetimes with 0.1% Tween20 in a Biotek ELx405 plate washer. The plates werethen read in a PerkinElmer TopCount platereader to measure the quantityof ³H-labeled peptide bound to the Flashplate surface, measured asdisintegrations per minute (dpm) or alternatively, referred to as countsper minute (cpm).

TABLE 7 Final concentrations of components for each assay variationbased upon EZH2 identity (wild-type or Y641 mutant EZH2) PRC2 EnzymeNon-radio- (denoted by EZH2 Peptide active SAM ³H-SAM identity) (nM)(nM) (nM) Wild-type 185 1800 150 Y641F 200 850 150 Y641N 200 850 150Y641H 200 1750 250 Y641S 200 1300 200 Y641C 200 3750 250

General Procedure for Wild-Type PRC2 Enzyme Assay on OligonucleosomeSubstrate. The assays was performed in a buffer consisting of 20 mMbicine (pH=7.6), 0.5 mM DTT, 0.005% BSG, 100 mM KCl and 0.002% Tween20,prepared on the day of use. Compounds in 100% DMSO (1 μL) were spottedinto polypropylene 384-well V-bottom plates (Greiner) using a Platemate2×3 outfitted with a 384-channel pipet head (Thermo). DMSO (1 μL) wasadded to columns 11, 12, 23, 24, rows A-H for the maximum signalcontrol, and SAH, a known product and inhibitor of PRC2 (1 μL) was addedto columns 11, 12, 23, 24, rows I-P for the minimum signal control. Acocktail (40 μL) containing the wild-type PRC2 enzyme and chickenerythrocyte oligonucleosome was added by Multidrop Combi (Thermo). Thecompounds were allowed to incubate with PRC2 for 30 min at 25° C., thena cocktail (10 μL) containing a mixture of non-radioactive and ³H-SAMwas added to initiate the reaction (final volume=51 μL). The finalconcentrations were as follows: wild-type PRC2 enzyme was 4 nM,non-radioactive SAM was 430 nM, ³H-SAM was 120 nM, chicken erythrocyteolignonucleosome was 120 nM, SAH in the minimum signal control wells was1 mM and the DMSO concentration was 1%. The assay was stopped by theaddition of non-radioactive SAM (10 μL) to a final concentration of 600μM, which dilutes the ³H-SAM to a level where its incorporation into thechicken erythrocyte olignonucleosome substrate is no longer detectable.50 μL of the reaction in the 384-well polypropylene plate was thentransferred to a 384-well Flashplate and the chicken erythrocytenucleosomes were immobilized to the surface of the plate, which was thenwashed three times with 0.1% Tween20 in a Biotek ELx405 plate washer.The plates were then read in a PerkinElmer TopCount platereader tomeasure the quantity of ³H-labeled chicken erythrocyte oligonucleosomebound to the Flashplate surface, measured as disintegrations per minute(dpm) or alternatively, referred to as counts per minute (cpm).

% Inhibition Calculation

${\%\mspace{14mu}{inh}} = {100 - {\left( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} \right) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{\left( {{Top} - {Bottom}} \right)}{1 + \left( \frac{X}{{IC}_{50}} \right)^{{Hill}\mspace{14mu}{Coefficient}}}}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

IC50 values for the PRC2 enzyme assays on peptide substrates arepresented in Table 3 below. In this table, “A” indicates IC50 values of10 μM and <50 μM; “B” indicates IC50 values of >3 μM and <10 μM; “C”indicates IC50 values of >1 μM and <3 μM; “D” indicates IC50 valuesof >0.3 μM and <1 μM; and “E” indicates IC50 values of <0.3 μM.

The compounds listed in Tables 1-6 were tested and the results weredescribed in Tables 8-13 below.

TABLE 8 Compound Number EZH2 IC50 A-1 E A-2 D A-3 D A-4 D A-5 D A-6 DA-7 E A-8 D A-9 D A-10 E A-11 D A-12 D A-13 E A-14 E A-15 E A-16 D A-17E A-18 E A-19 D A-20 D A-21 D A-22 D A-23 D A-24 D A-25 D A-26 D A-27 DA-28 D A-29 D A-30 D A-31 D A-32 D A-33 D A-34 D A-35 D A-36 D A-37 DA-38 D A-39 D A-40 D A-41 D A-42 D A-43 D A-44 D A-45 D A-46 D A-47 DA-48 C A-49 C A-50 C A-51 C A-52 C A-53 C A-54 C A-55 C A-56 C A-57 CA-58 C A-59 C A-60 C A-61 C A-62 C A-63 C A-64 C A-65 B A-66 C A-67 CA-68 B A-69 B A-70 B A-71 A A-72 A A-73 A A-74 A A-75 A A-76 A A-77 AA-78 A A-91 D A-92 C A-93 C A-94 B A-95 B A-96 B A-97 B A-98 A A-99 AA-100 A A-101 A A-102 A A-103 A A-104 A A-105 A A-106 A A-107 A A-108 AA-109 A A-110 A A-125 C A-126 D

TABLE 9 Compound Number EZH2 IC50 B-1 E B-2 D B-3 C B-4 E B-5 E B-6 DB-7 D B-8 E B-9 C B-10 D B-11 C B-12 C B-13 D B-14 D B-15 D B-16 E B-17C B-18 C B-19 D B-20 E B-21 D B-22 E B-23 D B-24 D B-25 E B-26 D B-27 DB-28 E B-29 E B-30 E B-31 E B-32 E B-33 E B-34 E B-35 E B-36 D B-37 DB-38 D B-39 D B-40 D B-41 D B-42 D B-43 D B-44 D B-45 D B-46 D B-47 DB-48 D B-49 D B-50 D B-51 D B-52 D B-53 D B-54 D B-55 D B-56 D B-57 DB-58 D B-59 D B-60 D B-61 D B-62 D B-63 D B-64 D B-65 D B-66 D B-67 DB-68 D B-69 D B-70 D B-71 D B-72 D B-73 D B-74 D B-75 D B-76 D B-77 DB-78 D B-79 D B-80 D B-81 D B-82 D B-83 D B-84 D B-85 D B-86 C B-87 CB-88 C B-89 C B-90 C B-91 C B-92 C B-93 C B-94 C B-95 C B-96 C B-97 CB-98 C B-99 C B-100 C B-101 C B-102 C B-103 C B-104 C B-105 C B-106 CB-107 C B-108 C B-109 C B-110 C B-111 C B-112 C B-113 C B-114 C B-115 CB-116 C B-117 C B-118 C B-119 C B-120 C B-121 C B-122 B B-123 B B-124 BB-125 B B-126 B B-127 B B-128 B B-129 B B-130 B B-131 B B-132 B B-133 BB-134 B B-135 B B-136 B B-137 B B-138 B B-139 B B-140 B B-141 B B-142 BB-143 B B-144 B B-145 A B-146 A B-147 A B-148 A B-151 A B-152 A B-153 AB-154 A B-155 A B-156 A B-164 B

TABLE 10 Compound Number EZH2 IC50 C-1 E C-2 E C-3 E C-4 E C-5 E C-6 EC-7 D C-8 D C-9 E C-10 E C-11 E C-12 E C-13 E C-14 E C-15 E C-16 E C-17E C-18 E C-19 E C-20 E C-21 E C-22 E C-23 E C-24 E C-25 E C-26 E C-27 EC-28 E C-29 E C-30 E C-31 E C-32 E C-33 E C-34 E C-35 E

TABLE 11 Compound Number EZH2 IC50 D-1 D D-2 E D-3 E D-4 E D-5 E D-6 ED-7 E D-8 D D-9 D D-10 E D-11 D D-12 E D-13 E D-14 E D-15 D D-16 E D-17E D-18 E D-19 D D-20 D D-21 D D-22 C D-23 E D-24 E D-25 E D-26 D D-27 DD-28 D D-29 D D-30 E D-31 D D-32 D D-33 E D-34 C D-35 C D-36 C D-37 ED-38 E D-39 E D-40 E D-41 E D-42 E D-43 E D-44 E D-45 E D-46 E D-47 ED-48 E D-49 E D-50 E D-51 E D-52 E D-53 D D-54 D D-55 D D-56 D D-57 DD-58 D D-59 D D-60 D D-61 D D-62 D D-63 D D-64 D D-65 D D-66 D D-67 DD-68 D D-69 C D-70 C D-71 C D-72 C D-73 C D-74 C D-75 C D-76 C D-77 CD-78 C D-79 C D-80 C D-81 B D-82 B D-83 B D-84 B D-85 B D-86 B D-87 BD-88 A D-89 A D-90 A D-91 D D-93 D D-94 D D-95 E D-96 E D-97 E D-98 D

TABLE 12 Compound Number EZH2 IC50 E-1 C E-2 E

TABLE 13 Compound Number EZH2 IC50 F-1 B F-2 D

INCORPORATION BY REFERENCE

The entire disclosure of each of the patent documents and scientificarticles referred to herein is incorporated by reference for allpurposes.

EQUIVALENTS

The invention can be embodied in other specific forms without departingfrom the spirit or essential characteristics thereof. The foregoingembodiments are therefore to be considered in all respects illustrativerather than limiting on the invention described herein. Scope of theinvention is thus indicated by the appended claims rather than by theforegoing description, and all changes that come within the meaning andrange of equivalency of the claims are intended to be embraced therein.

What is claimed is:
 1. A compound of Formula (lb) or a pharmaceuticallyacceptable salt thereof:

wherein: X₂ is N or CH; X₃ is CR₈; X₄ is C; Y₁ is CH; Y₃ is CR₁₁; R₆ isphenyl or a 5 to 6-membered heteroaryl selected from the groupconsisting of pyridinyl, pyrazolyl, pyrimidinyl, and furyl, wherein thephenyl and 5 to 6-membered heteroaryl are substituted with one or more-Q₂-T_(2,) in which Q₂ is a bond, and T₂ is unsubstituted 4 to7-membered heterocycloalkyl; R₇ is C₁-C₆ alkyl or cyclopentyl; each ofR₈ independently is H or methyl or ethyl; and R₁₁ is H.
 2. The compoundof claim 1, wherein R₆ is pyridinyl optionally substituted with one ormore -Q₂-T₂.
 3. The compound of claim 2, wherein X₂ is CH.
 4. Thecompound of claim 2, wherein T₂ is azetidinyl, oxetanyl, thietanyl,pyrrolidinyl, imidazolidinyl, pyrazolidinyl, oxazolidinyl,isoxazolidinyl, triazolidinyl, tetrahyrofuranyl, piperidinyl,1,2,3,6-tetrahydropyridinyl, piperazinyl, tetrahydro-2H-pyranyl,3,6-dihydro-2H-pyranyl, or morpholinyl.
 5. The compound of claim 4,wherein T₂ is piperazinyl.
 6. The compound of claim 5, wherein R₇ ismethyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl,s-pentyl, or n-hexyl.
 7. The compound of claim 6, wherein R₇ is s-butyl.8. The compound of claim 5, wherein R₁₁ is H.
 9. The compound of claim8, wherein R₈ is methyl.
 10. The compound of claim 9, wherein thecompound has R configuration.
 11. The compound of claim 9, wherein thecompound has S configuration.
 12. A compound of Formula (lb) or apharmaceutically acceptable salt thereof:

wherein: X₂ is CH; X₃ is CR₈; X₄ is C; Y₁ is CH; Y₃ is CR₁₁; R₆ ispyridinyl substituted with one or more -Q₂-T₂, in which Q₂ is a bond,and T₂ is piperazinyl; R₇ is s-butyl; R₈ is methyl; R₁₁ is H, andwherein the compound has S configuration.
 13. A pharmaceuticalcomposition comprising a compound of claim 1 or a pharmaceuticallyacceptable salt thereof and a pharmaceutically acceptable carrier.